The MC-2 fibrosarcoma, which is a transplantable tumour syngeneic for BALB/c mice, metastasizes to lymph nodes draining subcutaneous inoculation sites, and also to the lungs. T cell-mediated immunity was detected in Winn assays using spleens from excision immunized mice. T cell-mediated anti-tumour immunity was also detected in spleens from mice with small tumours but disappeared as the tumour burden increased. Protective immune spleen cell activity in the Winn assay was inhibited by prior addition of spleen cells from mice with large tumours, causing increased tumour incidence. Splenic metastases occasionally occurred in the MC-2 model, but were not demonstrable by bioassay in any of the experiments detecting splenic suppressor cell activity. In vivo protective activity was restored to advanced-stage tumour-bearer spleens by whole-body ionizing irradiation (0.5 and 2.5 Gy) of donor mice 15 h before sampling. Spleen cells from mice with small tumours remained protective after 1.5, 2.5 and 4.0 Gy of irradiation in vivo. These results are consistent with the properties of radiosensitive suppressor T cells. In contrast to reports in other tumour models, suppressor cells were not detected until late in the course of MC-2 development. This is surprising in view of the aggressively metastatic nature of MC-2. It is postulated that modulation of the anti-tumour immune response by the suppressor cells is associated with metastasis in this tumour model. The late appearance of both suppressor cells and metastatic cells in the spleen may reflect similar processes occurring earlier in regional lymph nodes.


The Walker 256 carcinosarcoma was shown to lack the enzyme 3-ketoacid CoA transferase. This suggests that ketone bodies cannot be used as a major substrate for the energy metabolism of this tumour. Systemic ketosis (1-2 mM acetoacetate plus 3-hydroxybutyrate) was induced both in tumour-bearing and in non-tumour-bearing rats with a diet containing 70% medium chain triglyceride. However, in rats bearing the Walker 256 tumour, this dietary ketosis did not reduce the tumour growth rate nor did it prevent the subsequent decrease in host body weight. Host body nitrogen losses were similarly unaffected. The ketosis induced in tumour bearing rats was shown to be abnormal since the blood glucose concentration of ketotic, tumour-bearing rats was significantly higher compared with that of ketotic non-tumour bearing rats . These results may partly explain why systemic ketosis failed to alter the growth and cachexia induced by the Walker 256 carcinosarcoma.


Photodynamic therapy consists of the systemic administration of a derivative of haematoporphyrin (Photofrin II) followed 24-72 h later by exposure of malignant lesions to photoradiation. We investigated the efficacy of this treatment after direct intratumoral injection of Photofrin II. This direct treatment regimen resulted in higher rates of inhibition of mitochondrial cytochrome c oxidase (5.13% J-1 cm-2 x 10(-1) and succinate dehydrogenase , respectively. A significant delay in tumour growth in vivo was observed in animals that received intratumoral injections 2 h before photoradiation compared to animals injected intraperitoneally at either 2 or 24 h before photoradiation. The treatment protocols were compared with control groups, consisting of Photofrin II administration intratumorally or intraperitoneally without photoradiation, or photoradiation in the absence of Photofrin II. These data indicate that the intratumoral injection regimen with Photofrin II enhanced the efficacy of photodynamic therapy. The greater delay in tumour growth observed after intratumoral administration of Photofrin II suggests a mechanism favouring direct cell damage.


The effect of dietary copper deficiency on tumour growth, neovascularisation and microvascular integrity was studied in the rat cremaster muscle. Male, weanling Sprague-Dawley rats were fed purified diets which were copper deficient (< 0.5 micrograms g-1 of diet) or copper adequate (5 micrograms g-1 of diet). Seven days after initiation of diets, a chondrosarcoma was implanted in the cremaster muscle of each rat. Five, 10 or 20 days after tumour implantation, rats were anesthetised and their cremasters prepared for observation by intravital microscopy. Intraarterial injection of fluorescein isothiocyanate-conjugated albumin and subsequent observation of fluorescence in the perivascular space indicated no difference in microvascular albumin leakage between the tumour vasculature of copper deficient and copper adequate rats. Neither tumour growth (assessed by wet weight), vascular density (assessed by light microscopy), nor any ultrastructural characteristics of the tumour or its vasculature (assessed by electron microscopy) were affected by copper deficiency. In view of findings by others which indicate changes in tumour characteristics with copper deficiency, we conclude that the copper dependency of tumour growth and vascularisation is a function of the type of tumour, the host tissue, or the conditions of copper depletion.


The radiosensitivity of human tumour cell lines and cells cultured from xenografts or biopsy specimens was measured using the adhesive tumour cell culture system (ATCCS). For cell lines the derived surviving fractions at 2 Gy were in good agreement with values obtained by clonogenic assay. However, the assay tended to overestimate survival at higher radiation doses, and thus to give a false impression of radioresistance. When cells taken from xenografts or tumour biopsies were cultured there was no evidence for selective growth of tumour cells: fibroblast-like cells commonly grew. Immunohistochemical staining against the intermediate filament, vimentin, supported the mesenchymal origin of the fibroblast-like cells. In cultures of artificial mixtures of tumour cells and fibroblasts, low proportions of fibroblasts were not excluded by the assay and consequently modified the radiation response. The majority of cultures grown from bladder carcinoma biopsy specimens appeared fibroblast-like, although in some cases clearly distinguishable colonies of tumour cells were also grown. In such tumour types the reliable measurement of radiosensitivity in cells taken from biopsies will require further development of techniques that allow the selective growth of tumour cells.


Six human melanoma xenograft lines grown s.c. in BALB/c-nu/nu mice were subjected to 31P-nuclear magnetic resonance (31P-NMR) spectroscopy in vivo. The following resonances were detected: phosphomonoesters (PME), inorganic phosphate (Pi), phosphodiesters (PDE), phosphocreatine (PCr) and nucleoside triphosphate gamma, alpha and beta . The main purpose of the work was to search for possible relationships between 31P-NMR resonance ratios and tumour pH on the one hand and blood supply per viable tumour cell on the other. The latter parameter was measured by using the 86Rb uptake method. Tumour bioenergetic status [the (PCr + NTP beta)/Pi resonance ratio], tumour pH and blood supply per viable tumour cell decreased with increasing tumour volume for five of the six xenograft lines. The decrease in tumour bioenergetic status was due to a decrease in the (PCr + NTP beta)/total resonance ratio as well as an increase in the Pi/total resonance ratio. The decrease in the (PCr + NTP beta)/total resonance ratio was mainly a consequence of a decrease in the PCr/total resonance ratio for two lines and mainly a consequence of a decrease in the NTP beta/total resonance ratio for three lines. The magnitude of the decrease in the (PCr + NTP beta)/total resonance ratio and the magnitude of the decrease in tumour pH were correlated to the magnitude of the decrease in blood supply per viable tumour cell. Tumour pH decreased with decreasing tumour bioenergetic status, and the magnitude of this decrease was larger for the tumour lines showing a high than for those showing a low blood supply per viable tumour cell. No correlations across the tumour lines were found between tumour pH and tumour bioenergetic status or any other resonance ratio on the one hand and blood supply per viable tumour cell on the other. The differences in the 31P-NMR spectrum between the tumour lines were probably caused by differences in the intrinsic biochemical properties of the tumour cells rather than by the differences in blood supply per viable tumour cell. Biochemical properties of particular importance included rate of respiration, glycolytic capacity and tolerance to hypoxic stress. On the other hand, tumour bioenergetic status and tumour pH were correlated to blood supply per viable tumour cell within individual tumour lines. These observations suggest that 31P-NMR spectroscopy may be developed to be a clinically useful method for monitoring tumour blood supply and parameters related to tumour blood supply during and after physiological intervention and tumour treatment. However, clinically useful parameters for prediction of tumour treatment resistance caused by insufficient blood supply can probably not be derived from a single 31P-NMR spectrum since correlations across tumour lines were not detected; additional information is needed.


A range of pure, monomeric porphyrins were synthesised and their localising capacities compared to HpD and Hp at 6 h and 24 h post injection in the mouse C6 intracerebral glioma model as well as in normal brain, skin, muscle, kidney, spleen, liver, lung and whole blood. The partition coefficients were examined between PBS and 2-octanol over the pH range 7.4-6.6 and pH profiles were established. A parabolic relationship was observed between log (porphyrin tumour concentration) at pH 7.4, with maximal tumour localisation at log (partition coefficient), pi, of approximately zero. Porphyrins with side chains with nett cationic character also exhibited up upward (parabolic) dependence on pi for most tissues studied, with maximal porphyrin localisation at pi of 0-0.5. In contrast, those porphyrins with nett anionic character exhibited a downward (negative) parabolic trend for all eight tissues studied, with minimal porphyrin localisation at pi of approximately zero. Four porphyrins  exhibited similar or better tumour localisation than HpD, and two (11 and 12) offer promise as lead compounds for the design of improved porphyrins for use in PDT.


A monoclonal antibody, H317, has been used for the sensitive and specific detection of placental-type alkaline phosphatase (PLAP) in sera, solubilized tissue extracts and fixed tumour tissue sections from patients representing a variety of ovarian tumours. PLAP was detected in over 30% of these sera and in most solubilized tumour tissue extracts. There was no association between circulating PLAP levels and either tissue extract levels or immunohistological staining of ovarian tumour tissue sections with H317. Nevertheless, immunohistology demonstrated the heterogeneity of cellular localization of PLAP within different tumours, and can often be of value in localizing tumour tissue.


From 1975 to 1988, 144 patients naive of treatment, with non-metastatic soft tissue sarcoma were treated at Fondation Bergonié by surgery, followed by radiotherapy and without chemotherapy. An analysis of prognostic variables was done on this population to determine patients for whom an adjuvant chemotherapy would be relevant. Prognostic variables in overall survival (OS), metastasis-free survival (MFS), disease-free and local free recurrence survivals were analysed by univariate and multivariate analysis. In multivariate analysis using Cox's model, only tumour depth and tumour grade were significant with the MFS end point, while tumour depth, tumour grade and tumour site were significant when considering OS. A predictive stratification for patients is proposed: a favourable prognostic group with grade 1 tumour or superficial, grade 2 tumour ; an intermediate prognostic group with deep, grade 2 tumour or superficial, grade 3 tumour ; and finally a poor prognostic group with deep, grade 3 tumour . Patients in the intermediate and poor prognostic groups who present a high metastatic risk are to be considered for adjuvant chemotherapy trials.


The covalent binding of a carotene moiety to one phenyl ring and meso-tetraphenyl-substituted porphyrins  efficiently quenches the photosensitising activity of the porphyrin while a relatively large yield of fluorescence emission around 650 nm is retained. Pharmacokinetic studies performed with two carotenoporphyrins (CPs) and the corresponding porphyrins (Ps) in Balb/c mice bearing an MS-2 fibrosarcoma show that the two Ps give a high selectivity of tumour localisation  and photosensitive tumour necrosis upon red light irradiation. For the same injected doses, the two CPs show no tumour-photosensitising activity even though they localise in the tumour in concentrations of the order of 10-40 micrograms g-1 at 24 h with tumour/peritumoral ratios larger than 10. Thus, the fluorescence emitted by these CPs in the tumour can be used for photodiagnostic purposes with no risk of skin photosensitisation. However, this approach is presently limited by the large accumulation and prolonged retention of the CPs in the liver and spleen.


The aim of this study was to determine the concentration of PDGF in vivo in neoplastic and non-neoplastic brain lesions. Fluid from cystic lesions and cerebrospinal fluid was tested in a radioreceptor assay that detects all described PDGF isoforms. High concentration of PDGF were found in cyst fluids from several astrocytomas, one metastatic melanoma, one metastatic lung adenocarcinoma and one intracerebral abscess. The PDGF concentrations were several times higher than the levels known to be required for maximal PDGF effects on cells in vitro. PDGF could also be detected in some non-neoplastic lesions, especially one intracerebral abscess. The finding of high amounts of PDGF in neoplastic lesions strongly supports the possibility that PDGF can be a mediator of tumour and stromal cell growth and motility in vivo. Comparison of PDGF and beta-thromboglobulin concentrations in the same fluids strongly indicates that the PDGF protein is locally produced rather than a result of platelet activation and derangement of the blood-brain barrier.


The use of a somatostatin analogue (SMS 201-995) has greatly facilitated the treatment of patients with the midgut carcinoid syndrome. Clinical studies have shown that SMS reduces the peripheral levels of tumour-produced serotonin (5-HT) and tachykinins, e.g. neuropeptide K (NPK), basally and after pentagastrin provocation. Some studies have indicated an inhibitory effect of SMS on tumour cell growth as well. In the present study we have investigated the effects of SMS on four different human midgut carcinoid tumours maintained in long term culture. Media levels of 5-HT and NPK-LI in tumour cell cultures decreased rapidly during incubation with SMS (10(-8)-10(-10) M) in all four tumours studied without evidence for tachyphylaxis (up to 6 weeks observation period). SMS treatment (10(-8) M) during 4 days reduced the media concentrations of 5-HT by 56%, while the intracellular contents of 5-HT were decreased by 27% indicating dual inhibitory effects on synthesis and secretion of 5-HT from tumour cells. The DNA contents of cultures were not affected by SMS (10(-8) M or 10(-10) M) treatment for 4 or 14 days. When tumour cell cultures were challenged with isoprenaline (IP) (10(-6) M) no reduction of the IP induced release of 5-HT could be detected after pretreatment of tumour cell cultures with SMS (10(-8) M) for 1 h, 4 h or 4 days. These studies provide evidence for a direct action of the somatostatin analogue on midgut carcinoid tumour cells, reducing both synthesis and secretion of hormones from tumour cells. This effect appears not to be related to inhibition of tumour cell growth. The inhibition of 5-HT secretion from tumour cells by SMS seems to operate via a second messenger system different from the one mediating the beta-adrenoceptor stimulated release of 5-HT.


The presence of T lymphocytes in solid tumours may reflect an ongoing immune response against the transformed cells. We have used polymerase chain reaction (PCR) technology to investigate the T-cell receptor variable-region gene (V-gene) usage in freshly isolated tumour-infiltrating lymphocytes (TILs) to look for a possible oligoclonality of T cells in the tumour area. We used 19 different V beta-family-specific primers. Peripheral blood lymphocytes and lamina propria lymphocytes from the same patients were also tested by PCR. Our results demonstrate a limited heterogeneity in the V-gene usage of TILs from seven patients with colorectal cancers, suggesting a local antigen-driven immune response at the tumour site.


In 31 patients with carcinoma of the breast the phenotype and activation status of tumour infiltrating lymphocytes (TILs) was analysed by flow cytometry. The predominant cells, in all patients, were T lymphocytes and in the majority of cases CD8+ (cytotoxic/suppressor) T lymphocytes were present in greater numbers than CD4+ (helper) T lymphocytes. There was no relationship between the degree of lymphocytic infiltration and either tumour stage or grade but there appeared to be an inverse correlation with the levels of oestrogen receptor (ER) in the tumour (P less than 0.01). Both populations of T cells had significantly higher numbers of cells carrying HLA DR (class II major histocompatibility antigen) than the equivalent populations in peripheral blood from the same patient group (P less than 0.001). The transferrin receptor was found on similar numbers of CD8+ T cells in peripheral blood and among the tumour infiltrating lymphocytes while more of the CD4+ T cells infiltrating the tumour were found to carry this receptor (P = 0.034). The Tac (CD 25) antigen was also on similar numbers of CD8+ T cells from both peripheral blood and the tumour but was on fewer of the CD4+ T cells in the tumour with respect to peripheral blood (P = 0.029). In both TILs and blood lymphocytes, the Tac antigen was consistently present on greater numbers of CD4+ T lymphocytes than on the CD8+ T lymphocytes (P less than 0.001) and as this is a component of the interleukin 2 (IL-2) receptor this may be of relevance to the use of IL-2 in TIL cancer therapy.


Using a one-stage kinetic chromogenic assay, we studied the procoagulant activity (PCA) of prostatic tissue in an experimental model of prostate cancer in the rat. PCA was present in homogenates of rat prostate glands containing either benign or malignant tumours. The procoagulant activated factor X directly and was provisionally characterised as a tissue factor-factor VIIa complex. There was no significant differences in PCA between control rats and rats exposed to carcinogens that did not develop tumour. Levels in rats that developed tumours were significantly higher (P < 0.01) than all other groups and there was a positive correlation between tumour weight and PCA . Furthermore, prostatic PCA levels were higher in the metastasis (P < 0.02). We conclude that PCA reflects the malignant phenotype in this animals, the PCA of the primary tumour was compared with that of the corresponding secondary deposit and levels were higher in the metastasis (P < 0.02). We conclude that PCA reflects the malignant phenotype in this model of experimental prostate cancer and suggest that this parameter is worth evaluating as a potential tumour marker in the human disease.


Photodynamic therapy (PDT) is an anti-cancer treatment which involves the systemic administration of a photosensitising drug which is preferentially absorbed by tumour tissue. Relatively little drug should be absorbed by the surrounding normal tissues. Tumour destruction is achieved when the tumour is illuminated with light of a wavelength which activates the photosensitising drug thereby inducing a cytotoxic reaction. However studies in many tissues have shown that the hoped for tumour selectivity is rarely achieved. Using the rabbit mandible and gingiva as our models we have studied the effects of various doses of PDT on the tissues of the oral cavity, namely mucosa, bone, muscle and salivary gland. The photosensitiser used was di-sulphonated aluminium phthalocyanine. Results show that whereas bone is extremely resistant to PDT the other tissues are vulnerable to it. In the case of muscle and salivary gland this susceptibility is very much dose related. In salivary tissue necrotising sialometaplasia was observed in areas of the gland adjacent to those that had undergone necrosis. All tissues were noted to heal or regenerate well following PDT injury.


Prostaglandins of the A series (PGAs) have been previously shown to inhibit the growth and to stimulate the differentiation of Friend erythroleukaemic cells (FLC) in vitro. In the present report we analysed the effect of PGA treatment in vitro on FLC tumorigenicity, and in vivo on FLC proliferation and on natural killer (NK) activity. PGA1 pretreatment of FLC in vitro for 5 days before inoculation into syngeneic mice slightly delayed tumour appearance, but did not significantly alter the pattern of tumour growth or mice survival, indicating that PGA1, at least in the conditions studied, did not affect FLC tumorigenicity. Daily treatment of mice with a long-acting synthetic analogue of PGA2  delayed tumour appearance, inhibited tumour growth, as measured by tumour weight and diameter, and increased the median mice survival time by 15-35%, depending on the schedule of treatment. Daily treatment with di-M-PGA2 strongly suppressed NK activity in normal mice but had no significant effect in tumour-bearing immunodepressed mice. PGA treatment of effector or target cells in vitro, or PGA added during the NK assay, had no effect on NK activity. We suggest that the chemotherapeutic effect of PGA is due to a direct action on tumour cell replication rather than to a stimulation of the host NK activity.


Cancer research has mainly focused on alterations of genes and proteins in cancer cells themselves that result in either gain-of-function in oncogenes or loss-of-function in tumour-suppressor genes. However, stromal variables within or around tumours, including blood and lymph vessels, stromal cells and various proteins, have also important impacts on tumour development and progression. It has been shown that disruption of stromal-epithelial interactions influences cellular proliferation, differentiation, death, motility, genomic integrity, angiogenesis, and other phenotypes in various tissues. Moreover, stromal variables are also critical to therapy in cancer patients. In this review, we mainly focus on the clinicopathological significance of stromal variables including angiogenesis, lymphangiogenesis, inflammatory infiltration, matrix metalloproteinase (MMP), and the particularly interesting new cysteine-histidine rich protein (PINCH) in colorectal cancer (CRC). The majority of cancer researchers have been focusing on tumour cells themselves and investigating their alterations in morphology, biology and function in tumour processes. However, stromal variables within or around tumours, including blood and lymph vessels, stromal cells, and various types of proteins, have not drawn enough attention even though they have important impacts on tumour development and progression. Tumour angiogenesis and lymphangiogenesis are the processes of creating new blood vessels or lymph vessels within and surrounding tumours. Stromal cells consist of various cell types such as infiltrating immune cells, fibroblasts, and endothelial cells. The extracellular matrix (ECM) is a complex structural entity surrounding tumour cells, and is often referred to as the connective tissue or ground substance. The ECM is composed of three major classes of biomolecules; structural proteins (collagen and elastin), specialized proteins , and proteoglycans ,2.A better understanding of the role of stromal variables in tumour development is required for designing appropriate therapeutic strategies against angiogenesis and stromal proteinases. A number of anti-angiogenesis elements and matrix metalloproteinase (MMP) inhibitors (MMPI) have recently been developed, and some have reached clinical trials -7. CompaIn this article, we mainly review the clinicopathological significance of stromal variables, including angiogenesis, lymphangiogenesis, inflammatory infiltration, MMPs and the particularly interesting new cysteine-histidine rich protein (PINCH) in colorectal cancer (CRC).In general, normal tissues have a barrier, preventing either endothelial cell migration or tumour cell invasion. The effect of this barrier can be interrupted by newly-formed stroma, namely stromatogenesis, during the process of tumour development . Stromat3, it must stimulate the host to creat its own vasculature to be able to continue growing. To accomplish this process, tumour cells induce adjacent blood vessels to sprout new vessels toward the tumour in a process called tumour angiogenesis [When a tumour grows larger than 1–2 mmogenesis . Since iogenesis . Lymphanogenesis -15. Lympogenesis .Both physiological and pathological stimuli such as hypoxia induce tumour cells, together with leukocytes, macrophages, mast cells and platelets, to secrete VEGF and other growth-related factors such as platelet-derived growth factor (PDGF) family proteins and their receptors (PDGFR), insulin-like growth factor (IGF) proteins and their receptors (IGFR), circulating endothelial precursor cell (CEPC), fibroblast growth factors (FGF)-2, angiopoietins, EphrinB2 and EphB4 . These fThe ligands of the VEGF family include VEGF-A, VEGF-B, VEGF-C, VEGF-D and VEGF-E. All five ligands have different roles in the process of angiogenesis and lymphangiogenesis. Each of the VEGF-family ligands binds to one or more of three known VEGF receptors (VEGFR): VEGFR-1 , VEGFR-2 (Flk-1 or KDR) and VEGFR-3 (Flt-4). VEGFR-1 organises blood vessels, and has a high affinity for VEGF-A and VEGF-B. VEGFR-2 activates blood vessel proliferation by binding to VEGF-A, VEGF-C, VEGF-D, and VEGF-E. VEGFR-2 is expressed in lymphatic endothelial cells ,16-18. VIt has been found that high microvessel density (MVD) is associated with VEGF -27 and Vet al. have analysed a microvessel pericyte coverage index (an index of microvessel maturation) in relation to clinicopathological significance in CRC [et al. have applied the anti-α-smooth muscle actin (SMA) marker to determine the microvessel pericyte coverage index [Many research groups have studied CRC and shown that a high grade of MVD is related to a larger tumour size ,35,36, ne in CRC . The resge index . There age index ,45. Whilge index .There are a few conflicting reports in CRC regarding the clinicopathological significance of MVD, in which MVD is not associated with tumour stage ,43,47, vet al. observed an increased vascularization of metastases in the liver after resection of the primary CRC [There are even opposite results from two studies in which higher values of MVD appear to be in the early stages of CRC , and cormary CRC . This remary CRC .Hypercoagulation in cancer patients is another factor for tumour progression. Substantial evidence from preclinical experiments and clinical practice has supported the association between activation of blood coagulation and progression of the cancer. Cancer patients display a wide range of coagulation disorders from asymptomatic laboratory changes to massive thromboembolism and disseminated intravascular coagulation. About 50% of all cancer patients and 90% of patients with metastasis have abnormalities in coagulation tests. CRC is the second most common cancer diagnosed in patients with thromboembolic events. Blood vessel thrombosis leads to impairment of blood flow, ischemia, and organ damage. The hemostatic complications are the second most common cause of death in cancer patients .Studies have shown that plasma D-dimer levels, representing activation of coagulation and fibrinolysis, are increased in most patients with CRC compared to patients with benign colorectal disease. Furthermore, the D-dimer level is positively related to tumour size, wall penetration, lymph node invasion, and hepatic metastasis . In a muAlthough the mechanism behind hypercoagulation in cancer patients is unclear, the main factor responsible for hypercoagulation has been considered to be cancer itself. It has been shown that tumour cells activate the coagulation system by producing and secreting procoagulant/fibrinolytic substances and inflammatory cytokines, as well as physically interacting with blood  or vascular cells . This acA recent study shows that VEGF is highly expressed in primary CRC compared to the corresponding adjacent normal mucosa . VEGF exIncreased expression of VEGF-A in CRC is associated with lymphatic metastases . IncreasThere are few studies of VEGFR expression in CRC. Some studies show that either VEGFR-2 or -3 expression on CRC does not differ from that in the normal mucous of the colorectum ,22, whilThere are controversial results regarding the role of VEGF and VEGFR in CRC. For example, levels of VEGF expression in primary CRC and liver metastases do not significantly differ . VEGF-A et al. examined the expression patterns of several VEGF-A isoforms in 228 established xenografts originating from various human solid tumours including colon cancer. VEGF121/VEGF165 were seen in 27 xenografts and VEGF121/VEGF165/VEGF189 in 201 xenografts. VEGF189 is more frequently expressed in all tumour xenografts than in primary tumours, indicating that VEGF189 contributes to the successful xenotransplantability of various solid tumours through the induction of stromal vascularization [Even splicing variants in certain members of the VEGF family play different roles in tumour development. For example, the VEGF-A gene, located on chromosome 6p21.3 with eight exons, gives rise to several distinct isoforms of VEGF-A through alternative mRNA splicing. The more common isoforms of human VEGF-A consist of VEGF121, VEGF145, VEGF165, VEGF165b, VEGF189, and VEGF206, and other isoforms such as VEGF148, VEGF162 and VEGF183 have also been reported. VEGF-B also has different isoforms such as VEGF167 and VEGF186 ,71. Thesrization . Althougrization ,75.et al. observed that intratumoural lymphatic vessels were present in the majority of colon carcinomas (91%), and had a significantly higher density in the submucosa near the tumour [There are only a few studies of lymphatic density in CRC. Parr and Jiang examined lymph vessel status by using several lymphangiogenic markers , and found that their expression was higher in CRC compared to normal mucosa . In adene tumour . Furthere tumour .Like new blood vessels, lymphatics at the centre of tumours do not function as well in spreading tumour cells as they do at the invasive margin of tumour. The intratumoural lymph vessels are often compressed and smaller, while the lymph vessels around tumours are often enlarged and hyperplastic. These enlarged vessels may collect tumour cells from the tumour and possibly contribute to lymphatic metastasis. VEGF-C and VEGF-D induce not only the density but also the enlargement of lymphatic vessels, which leads to metastases to the regional lymph nodes -80.Anginogenesis, lymphangiogenesis, VEGF, and VEGFR are increased in CRC, compared to normal mucosa and adenoma, and are further related to more malignant features of CRC including poorer survival.There are two types of immune responses, innate and adaptive immunity. Innate immunity reacts rapidly to molecular patterns found in microbes, independent of prior contact with a pathogen. The adaptive immune response is specific and has immunologic memory . Immune The TII response may have dual effects in the development and progression of the tumour. On one hand, inflammatory cells can kill tumour cells, resulting in tumour regression and a greater chance of survival for the cancer patient. On the other hand, production of cytokines and growth factors derived from TII can stimulate tumour cells to grow and emigrate. The effects on tumour development may depend on host- and tumour-specific features such as the immunoresponse of the host or the type and biological features of the tumour -93. For Many studies have shown that a high grade of lymphocytic infiltration or TII in CRC is related to favourable survival of patients -106, andet al. have analysed 60 slides from 60 tissue blocks from 30 colonic carcinomas, and the slides are circulated twice to six histopathologists with varying experience [Notably, TII in the inner part of the tumour is not significantly related to clinicopathological variables including patient survival, but abundant TII at the invasive margin of the tumour predicts a favourable prognosis in CRC patients . These rperience . Five ouTAMs are highly localized at the invasive margin of CRC compared with the central area of the tumour ,122,123.It has been shown that TAMs are positively associated with angiogenesis in CRC . Such anPatients with CRC present reduced numbers of peripheral blood DCs compared with healthy controls . Furtheret al., using markers CD1a and CD208 for DCs in CRC, found that patients with high levels of either CD1a- or CD208-positive DCs had shorter survival [A study in 170 patients with rectal cancer using CD1a as a marker for DCs did not find that DCs are related to survival . Notablysurvival . Howeversurvival . Since tsurvival . The disTII in CRC, especially at the invasive margin of tumours, plays a critical role against tumour development and aggressiveness, based on the relationship of strong TII with better differentiation, earlier stage, lower rates of local/distant recurrence and better survival.Cell adhesion to the ECM is mediated by integrins. Focal adhesion (FA) is an integrin-rich cell adhesion sites, containing cytoskeletal signalling molecules including FA kinase, integrin-linked kinase (ILK), talin, vinculin and paxillin. ILK is an intracellular serine/threonine protein kinase regulating integrin-mediated cell adhesion, E-cadherin expression, pericellular fibronectin matrix assembly and cellular proliferation and survival -142. ThrDuring tumour progression, tumour cells must remodel the matrix either by expressing or degrading ECM proteins to facilitate communication and escape control by the microenvironment. The remodelling of the microenvironment surrounding tumour cells leads to the release of ECM-associated growth factors which may function to suppress or induce tumour growth . Thus, mMMPs are a family of ECM degrading proteinases, secreted by both tumour and stromal cells. Based on substrate specificities and sequence characteristics, the classic MMP family members can be divided into at least four subgroups; collagenases, gelatinases, stromelysins, and matrilysins. So far, 23 different MMPs, MMP-1, -2, -3, -5, -7, -8, -9, -10, -11, -12, -13, -14, -15, -16, -17, -18, -19, -20, -21, -23, -25, -26, and -28, and four tissue inhibitors of metalloproteinases (TIMPs), TIMP-1, -2, -3 and -4, have been cloned -147. MMPUnlike classical oncogenes, MMPs are not upregulated by gene amplification or activating mutations, and the increased MMP expression in tumours is probably due to transcriptional changes. This might be the result of activation of oncogenes or loss of tumour suppressors. It has been demonstrated that MMP7 is upregulated by the transcription factor PEA3, and MMP1 and MMP13 are downregulated by the tumour suppressor p53 -154. Theet al. have reported that expression of MMP-3 and -13 is greater in CRC but MMP-1 expression is not [et al. have observed strong expression of MMP-3 and -10 in colon cancers but not MMP-13 [Among MMPs, MMP-1, -2, -3, -7, -9, -11, and -13 have been studied the most. Expression of MMPs, such as MMP-1, -2, -3, -7, and -9, is greater in CRC than in normal mucosa or in adenoma of the colon/rectum -167. Hign is not . Bodey et MMP-13 . Unlike t MMP-13 .et al. have found that hereditary nonpolyposis colorectal cancer syndrome (HNPCC) exhibited a significantly lower expression of MMP-1 and -9, whereas sporadic CRCs usually have increased expression of MMPs. These findings on the basis of lower matrix-degrading properties of the fibroblastic tumour stroma in HNPCC may help us to understand why HNPCC, compared with sporadic CRC, has lower malignancy, for example, a better prognosis in HNPCC patients [Regarding the clinicopathological significance of MMPs, most findings have shown that increased expression of MMPs is related to more malignant features of CRCs. MMP-1 correlates with poor differentiation , advancepatients .During tumour invasion and metastasis, tumour cells must pass a series of basement membrane and ECM barriers. The basement membrane is the first and most important barrier for tumour cells to penetrate to complete invasive and metastatic processes. The ECM must be broken down to permit tumour cells to invade surrounding tissues or metastasise to other organs. Furthermore, some MMPs stimulate angiogenesis for promoting tumour growth and invasion. Although the mechanism by which MMPs enhance the invasive and metastatic competence of tumour cells seems straightforward, the specific role of distinct MMPs in the progression of tumour invasion and metastasis is more complex than has been assumed. For example, MMP-2 does not show an association with tumour differentiation, stage, metastasis, or patients' prognosis . OppositWe found that ST3 (MMP11) is positively related to PINCH expression in CRC (unpublished data), and, both ST3 and PINCH are present in stromal fibroblasts around tumour cells but not in tumour cells themselves ,179. TheST3-null mice have an increased frequency of apoptosis and necrosis compared to the wild-type hosts [Although the exact mechanism of ST3 action is unknown in tumour development, one hypothesis is that ST3 is implicated in basement membrane remodelling through release or activation of growth factors or cytokines stored in the ECM. ST3 degrades insulin-like growth factor-binding protein-1 (IGFBP-1), leading to cellular proliferation and survival . It has pe hosts . ST3 inhpe hosts . ST3 is pe hosts , but pospe hosts . Thus, SST3 protein is present in the stromal fibroblasts around tumour cells but not in tumour cells themselves. ST3 expression, determined by Northern blot, in-situ hybridisation or immunohistochemistry, is undetectable in normal colorectal mucosa, detectable in low levels in adenoma, and at higher levels in primary CRC ,189-194 The rate of extensive ST3 expression is significantly higher in the de novo group than in the ex adenoma group. Histopathologically, the de novo group has a significantly higher rate of cases with an infiltrative invasion pattern. These results may indicate that ST3 expression is implicated in a greater invasive potential of CRC . As showThe MMP family is likely to be involved in early development of CRC via remodelling of the basement membrane, inhibition of tumour cell apoptosis and the host immune response, and angiogenesis activation. MMPs also play critical roles in tumour aggressiveness based on their relationships with advanced tumour stage, high frequency of recurrence and worse prognosis. However, MMPs can also suppress tumour growth, invasion and metastasis.PINCH was originally identified by Rearden in 1994 from screening a human cDNA library with antibodies recognizing senescent erythrocytes . The PINPINCH interacts directly with ILK through its LIM1 domain binding to the first of four ankyrin (ANK) repeat domains at the ILK N-terminus. The C-terminal domain of ILK has certain homologies with the catalytic domains of serine/threonine protein kinases. This kinase-like domain is able to interact with several components of cell-matrix contact sites including CH-ILKBP , β1, β2, and β3 integrin cytoplasm tails, β-parvin (affixin), and paxillin -207. PINPINCH can also bind to Nck2, an additional PINCH binding partner, through the LIM4 domain of PINCH and the SH3 domain 3 of Nck2. Nck2, as an SH2/SH3 adaptor protein, is an important component of the signalling pathways of growth factor receptors including epidermal growth factor (EGF) and PDGF receptors, and can modulate actin dynamics by interacting with p21-activated kinase. Therefore, PINCH, by mediating the formation of the complex between ILK and Nck-2, is involved in the regulation of ILK function, which is implicated in many critical physiological and pathological processes ,209-211.et al. identified a second member of the PINCH family, PINCH2, and therefore, PINCH was renamed to PINCH1 [Zhang o PINCH1 . The PINo PINCH1 .PINCH2 also localizes to cell-ECM adhesion sites but only the LIM1 domain binds to ILK, suggesting that PINCH2 may potentially interact with other components of the cell-ECM adhesion structure. In addition to regulating the PINCH1-ILK interaction, cell spreading and migration, PINCH2 may participate in the regulation of nuclear processes since PINCH2 is present at high levels in the nucleus of the cell . HoweverAfter identifying the PINCH gene, Rearden's group in 2002 further analysed PINCH protein expression determined by immunohistochemistry using a polyclonal antibody against PINCH in human tissues, and observed that PINCH expression was markedly upregulated in the tumour-associated stroma of many common cancers including breast, prostate, lung, skin, and colon cancers, compared to the corresponding normal tissues . As showThe intensity of PINCH expression in the inner part of primary tumour is not significantly related to survival and other clinicopathological variables including Dukes' stage, growth pattern and grade of differentiation . LocalizThe PINCH protein is detected not only in fibroblasts, but also in myofibroblasts and in a proportion of endothelial cells of the tumour vasculature  supporting the involvement of PINCH in promoting tumour-stromal interactions that support tumour progression ,212. PINThe adapter protein PINCH, a new component of the cell-ECM adhesion structure, may have an important role in tumour invasion and metastasis, via tumour-stromal interactions, resulting in a poor prognosis. It is of interest to further study the biological and clinicopathological significance of PINCH 1 and PINCH2 in tumours.Although conventional chemotherapy and radiotherapy have improved the outcome for CRC patients, the benefits of the treatments are still under investigation, especially in patients with advanced-stage tumours. The side effects of the treatments and resistance to the treatments are still major problems. Because of a lack of specific markers to select patients for suitable treatments many patients have been overtreated by chemotherapy and radiotherapy. Thus, biomarkers for both tumour cells and stroma are urgently needed to complement current tumour stage in terms of response to treatments. Compared to tumour cells, the stroma has been shown to be a more attractive therapeutic target. Firstly, regulation of stromal activity could affect tumourigenesis in different ways including inhibition of angiogenesis, lymphangiogenesis and MMP activity as well as activation of certain TII, to stabilize and regress the primary tumour. Secondly, some stromal factors, such as endothelial cells, are highly accessible to circulating drugs or drug carriers . ThirdlyAntiangiogenic therapy is a new promising strategy for inhibiting tumour growth, development and metastasis. A number of potential angiogenic inhibitors have been developed to affect endothelial cell proliferation, migration and survival, and some of these agents have entered clinical trials in CRC patients. PTK/ZK acts on VEGFR-1, -2 and -3, SU 11248 on VEGFR-1, -2, -3 and PDGFR, ZD 6474 on VEGFR-1, -2, -3 and EGFR, BAY 43–9006 on VEGFR-2, -3 and PDGFR, AEE 788 on VEGFR-1, -2 and EGFR, Imatinib on PDGFR, and gefitinib and erlotinib on EGFR . Among tImmunotherapies, as less toxic treatment modalities, have emerged as potentially attractive alternatives for cancer therapy. It has been shown that when patients with resectable recurrent CRC are treated with interleukin-2 before surgery, there is an increase of eosinophilic infiltration in tumour tissue, indicating that interleukin-2 increases the host response to the tumour . In anotMMPIs, such as BAY12–9566, AG3340, BMS275291 and CGS27023A/MPI270, have been shown to inhibit tumour growth in preclinical models. However, this treatment has achieved minimal success in patients with advanced cancers in clinical trials. The differences between preclinical and clinical results with MMPIs are not just due to differences between animal and man but are rather related to the stage of disease, different endpoints, and the treatment methods. For example, MMPIs are unlikely to be effective in patients with advanced-stage cancer. However, there are significant preclinical data to support a role for MMPIs in earlier stages of cancer. In addition, MMPIs are used to treat patients with endpoints of increased time to progression or improved outcome. In comparison, preclinical experiments performed in animals is to examine tumour progression with the final endpoints of reduced tumour number and size or metastasis. Finally, patients are given the maximum tolerated dose and this is often limited by musculoskeletal side-effects while animal studies used escalating doses of MMPIs that are not limited since mice are less susceptible to this side-effect. Thus, it is of importance to design MMPIs for selected patients. It is also important to select MMPIs-1, -3, -7, -9, and -13 as potential agents, since the expression of the corresponding MMPs correlates with metastasis and poor prognosis in cancer patients. Moreover, MMPs have a dual function in tumour development; namely, they play critical roles not only in tumour aggressiveness but also in suppression of tumour growth. This complicates treatment targeting MMPs. Based on accumulated data, it is recommended that future MMPI trials focus on: (1) patients with early stage cancer; (2) the use of MMPIs along with chemotherapy; (3) the measurement of MMPs in tumour tissue and blood as a means of identifying patients who are more likely to respond to MMPI therapy; and (4) identification of biomarkers that reflect activation or inhibition of MMPs in vivo .et al. examined patients with CRC at stage II or III, who underwent potentially curative resection. The patients were divided into two groups: one group received postoperative administration of fluoropyrimidines (such as UFT and 5'-DFUR), and the other group underwent surgery alone. The disease-free survival rate in the chemotherapy group was significantly higher than that of the surgery-alone group. However, this difference was not seen between the two groups who had MMP-9 positive tumour. Thus, the efficacy of the chemotherapy may not be great for patients with a tumour positive for MMP-9 [MMPs have been suggested as biomarkers for selecting cancer patients for suitable treatments. Ogata or MMP-9 .Although there are promising results in the treatment of cancer patients with immunotherapy, anti-angiogenesis and MMPIs, the roles and effects of these treatments are still under investigation. An important challenge is how to combine biological agents with different cytotoxic agents in CRC. Another important challenge is to combine therapeutic targets of tumour cells with those of stromal factors.It has been suggested that CRC is caused by both environment, including life style, and genetic predisposition. In general, the number of genetic alterations is increased from the first genetic change in a normal endothelial cell to several genetic alterations in the late stages of cancer cells. Moreover, in the tumour stroma, the number and construction of blood and lymph vessels are altered by stimulation with many stromal factors such as VEGF, VEGFR, TII, MMP, PINCH, and others. Interactions between these growth and anti-growth stromal factors in the tumour stroma affect the formation, development and progression of CRC Figure . AnticanCRC: colorectal cancer. DC: dendritic cell. ECM: extracellular matrix. FA: focal adhesion. HNPCC: hereditary nonpolyposis colorectal cancer syndrome. IGF: insulin-like growth factor. ILK: integrin-linked kinase. MMP: matrix metalloproteinase. MMPI: Matrix metalloproteinase inhibitor. MVD: microvessel density. NK: natural killer. PDGF: platelet-derived growth factor. PINCH: particularly interesting new cysteine-histidine rich protein. SMA: smooth muscle actin. ST3: stromelysin-3. TAM: tumour-associated macrophage. TII: tumour inflammatory infiltration. TIMP: tissue inhibitors of metalloproteinases. VEGF: vascular endothelial growth factor. VEGFR: vascular endothelial growth factor receptor.


The murine monoclonal antibody H17E2 recognises placental alkaline phosphatase (PLAP), an antigen present in the human term placenta and also expressed by many tumours. The antibody is of value in both immunoscintigraphy and radioimmunotherapy in testicular and ovarian cancer. The small size of genetically engineered single chain antibodies (SCAs) should give diagnostic and therapeutic advantages of improved tumour penetration and increased blood clearance compared to IgG. Employing recombinant DNA techniques a SCA based on H17E2 has been expressed in Escherichia coli and has been shown to bind placental alkaline phosphatase specifically. When administered to nude mice bearing human tumour xenografts, the H17E2 SCA effectively localised to tumour whilst a co-administered non-specific SCA did not. H17E2 SCA achieves tumour: blood ratios that are superior to those achieved with whole IgG, probably owing to its rapid blood clearance. We conclude that the H17E2 SCA is suitable for further investigation as an agent for clinical imaging and therapy. Additionally, the SCA can also be used for the construction of antibody based fusion proteins to target other effector functions to tumour cells.


We have examined a series of 13 benign and 27 malignant human gliomas for evidence of molecular abnormalities of proto-oncogene and putative tumour suppressor gene loci. The results indicated that specific molecular lesions were associated with increasing grades of malignancy. Thus, loss of genetic material on chromosome 17 was present with approximately equal frequency in both benign and malignant gliomas, whereas loss of loci on chromosome 10 was seen only in malignant gliomas. Only the most malignant tumours, known as glioblastoma multiforme, had more than one molecular abnormality in the same tumour. These findings may contribute to our understanding of glial tumour development, as well as improve the accuracy of tumour diagnosis.


Limonene, a monocyclic monoterpene, occurs naturally in orange peel oil. It has been shown to exhibit both chemopreventive and chemotherapeutic activity without toxicity in rodent models. In this study we examined the effect of limonene both at maximally optimal and suboptimal doses and in combination with suboptimal doses of 4-hydroxyandrostrenedione on nitrosmethylurea-induced rat mammary tumours. A 10% limonene dose mixed in the diet caused tumour regression in all animals. A 5% limonene dose was only able to cause regression in 50% of the rats (P < 0.05). A suboptimal dose of 4-hydroxyandrostrenedione (12.5 mg kg-1) resulted in tumour regression in 75% of rats. A combination of 5% limonene with 4-hydroxyandrostrenedione (12.5 mg kg-1) resulted in a greater tumour regression (83.3%) than either agent given individually (P < 0.001 and 0.006 for limonene/4-hydroxyandrostrenedione vs limonene alone and 4-hydroxyandrostrenedione alone respectively.


Dose limiting systemic toxicity prevents sufficient exploitation of the steep dose response relationship of most anticancer agents. In our rat liver tumour model , isolated liver perfusion allows administration of higher doses of mitomycin C than hepatic artery infusion, while systemic toxicity remains minimal. To determine the temporal pattern of mitomycin C induced cytokinetic changes, we analysed flow cytometric DNA histograms of CC531 liver tumours from rats treated with high dose mitomycin C (3.2 mg kg-1) via hepatic artery infusion and sacrificed at different time intervals after treatment. Between 12 and 36 h after treatment, the fraction of cells in late S and G2/M phase had markedly increased. The effects of administration of the respective maximally tolerated doses of mitomycin C in isolated liver perfusion and via hepatic artery infusion on progression of tumour cells through the cell cycle and on gross tumour growth were compared. Isolated liver perfusion with mitomycin C resulted in a significant increase in the proportion of cells in mid and late S, and in some accumulation of cells in early S and G2/M phase at 24 and 48 h after treatment. In contrast, after hepatic artery infusion a significant increase of the fraction of cells in G2/M phase was observed at 24 h after treatment. Monitoring tumour growth after isolated liver perfusion five out of seven rats showed a complete tumour remission, while after hepatic artery infusion only a minimal growth delay was detected. This study demonstrates that isolated liver perfusion in the rat CC531 liver tumour model allows the administration of a well-tolerated dose of mitomycin C being high enough to induce a marked DNA synthesis inhibition and even complete tumour remission.


The growth rate of the MAC16 tumour in cachectic animals was significantly enhanced by the hypolipidemic agent bezafibrate, while the growth rate of a histologically similar tumour, the MAC13, which grows without an effect on host body compartments was unaffected. Growth of the MAC16 in vitro was unaffected by bezafibrate, suggesting that it was an in vivo phenomenon only. The stimulatory effect of bezafibrate correlated with the maximum plasma levels of free fatty acids (FFA) arising from the catabolism of adipose tissue. Accumulation of 14C-lipid from 1-14C-triolein administered by intragastric intubation was enhanced in heart, gastrocnemius muscle and tumour of bezafibrate treated animals, while the total lipid absorption did not differ from solvent treated controls. The increased lipid accumulation in the heart, but not the tumour correlated with an increased tissue lipoprotein lipase level. The increased tumour level may arise from an increased uptake of FFA arising from a weakening of the bonds between FFA and albumin. These results suggest that growth of certain tumours is dependent on maintaining sufficient lipid levels and that the lipid mobilising effect of the tumour may be necessary to sustain tumour growth.


In order to assess the time at which the distant metastases were initiated, a model has been developed to simulate the natural history of human breast cancer. The metastasis appearance curves were fitted to those observed for tumours of various sizes among the 2648 patients treated at the Institut Gustave Roussy from 1954 to 1972. The model assumes that metastases are initiated when the tumour reaches a threshold volume (distribution of this volume was estimated in a previous article). Two patterns of growth were considered: exponential and Gompertzian. Distributions of tumour and metastases doubling times are fixed according to the literature. A relationship between tumour and metastasis doubling time is estimated. Simulations were used to optimize metastases growth duration as a function of the metastasis doubling time. The ages of the metastases at tumour diagnosis are calculated. With exponential growth, it was necessary to introduce correlations to obtain a satisfactory fit of the metastases appearance curves: between the tumour volume at diagnosis and the doubling time (R1 = -0.3), and between the tumour volume at metastasis initiation and the doubling time (R2 = 0.3). The growth duration of the metastases before their detection was found to equal about 18 metastases doubling times at detection and the mean ratio between the doubling time of a tumour and its metastases equal to 2.2. With Gompertzian growth, it was impossible to adjust satisfactorily the proportions of metastases at diagnosis as a function of the primary tumour volume. However, when we ignore this, the best fit was obtained when the duration of metastases growth before detection was about the same as for exponential growth. With either growth pattern, the model predicts that the proportion of patients with metastases would be reduced by approximately 30% if the primary tumours were treated 12 months earlier. This prediction is consistent with the results of the screening programs for breast cancer.


To investigate the influence of culture conditions on the in vitro responses of tumour cells to anticancer drugs, the sensitivities observed with the soft agar methods of Hamburger & Salmon (1977) (H-S) and of Courtenay & Mills (1978) (C-M) were compared. In all cases the ID50 values were determined from dose-response curves. Six human tumour cell lines exposed to 10 different agents, and 9 patients' melanomas exposed to 5 different agents, were examined. In the studies of cell lines the H-S method gave higher sensitivity values than the C-M method in 38 out of 52 cases, whereas in 14 cases the results were the same. In the patients' tumours the H-S method gave higher sensitivity in 21 of 35 cases, equal sensitivity in 11, and lower sensitivity in 3 cases. In many instances the ID50 values obtained with the two test systems differed by factors of 10 or more, both in the case of cell lines and tumour specimens. Systematic alterations in the culture conditions indicated that the presence or absence of rat erythrocytes is the most important factor responsible for the differences observed. Also, other factors, such as supplements (in the H-S method) and the use of different serum types, appeared to influence both colony growth and chemosensitivity.


We assess the feasibility of using the MTT assay as a measure of cell viability in chemosensitivity testing in ovarian malignancy. The assay utilises the conversion of the tetrazolium salt MTT to formazan by dehydrogenase enzymes in living cells. We show that the optical density of the formazan produced from MTT is directly proportional to the number of live cells tested. Optimum MTT conversion occurred after 4 h incubation and dimethyl sulphoxide was found to be the most suitable solvent for the formazan. Seventy-five samples of ascitic fluid and/or solid tumour were collected from 56 patients with FIGO stage III-IV ovarian adenocarcinoma. Malignant cell suspensions with a viability greater than 75% were prepared from 95% of ascitic fluid and 75% of biopsy samples by simple techniques. The effect of cytotoxic drugs was assessed in 91% of patients included in the study. Variation in drug effect between patients was evident following a 48 h incubation period and was reproducible. Overall platinum and anthraquinone analogues produced the greater effect but resistance did occur. Our results mirrored reported clinical response rates. Only one sample tested against chlorambucil showed any drug effect. As this assay produces results in a high percentage of tests and is rapid and simple it appears suitable for prospective clinical trials to correlate the in vitro results with in vivo response.


Urinary recovery of enterally administered probes is used as a clinical test of intestinal mucosal permeability. Recently, evidence has been provided that the recovery of some but not all sugar probes is dependent on the amount of diuresis and renal function. The aim of this study was to assess the effect of fluid loading on the urinary recovery of sugar probes in healthy volunteers.O-methyl-D-glucose (3-OMG), 0.5 g of D-xylose, 1.0 g of L-rhamnose, and 5.0 g of lactulose on two different days. The volunteers were randomized to receive either 2 litres of Ringer acetate or no fluid during the following 3 hours. The sugar concentrations were measured in 5-hour urine samples period.In a cross-over study, 10 healthy volunteers ingested 100 ml of a solution containing 0.2 g of 3-Fluid loading increased urine production and urinary recovery of xylose. Fluid loading did not influence the urinary recovery of 3-OMG, L-rhamnose, or lactulose. Neither the lactulose/rhamnose ratio nor the 3-OMG/rhamnose ratio changed.Fluid loading increases mediated carbohydrate transport but not the lactulose/rhamnose ratio, after oral sugar administration in healthy volunteers. It remains to be determined whether sugar probes are handled differently in response to fluids in patients with organ dysfunctions. Mucosal permeability is one of the few functions of the gastro-intestinal tract that can be quantified in the clinical setting. Increased intestinal permeability in critically ill patients has been associated with endotoxemia and remote organ failure . MucosalIn experimental animals, fluid loading increased the urinary recovery of intravenously administered lactulose but not rhamnose, and thereby increased the lactulose/rhamnose (L/R) ratio , suggestO-methyl-D-glucose (3-OMG), 0.5 g of D-xylose, 1.0 g of L-rhamnose, and 5.0 g of lactulose on two different days  with a median age of 32 years (range 23 to 42 years) were studied on the morning after an overnight fast. The subjects were not allowed to consume alcoholic beverages or to use drugs the day before the testing days. The study was approved by the Ethics Committee of Kuopio University Hospital. The research was performed in accordance with the Declaration of Helsinki. All volunteers gave written informed consent to participate in the study. In a cross-over design, the volunteers ingested 100 ml of a solution containing 0.2 g of 3-The data were analysed with Wilcoxon signed-rank test. Data are presented as medians and ranges.Fluid loading increased urine production during the 5-hour collection period Table , Fig. 2.Altered intestinal permeability has frequently been reported in patients requiring intensive care after major surgery and in sepsis. The mechanisms of these alterations are not entirely defined. In addition to permeability of intestinal epithelium, both pre-mucosal and postmucosal factors can affect the urinary excretion of enterally administered sugar probes ,8. FluidWe studied the effect of a postmucosal factor, fluid loading, on the urinary excretion of enterally administered sugar probes in healthy volunteers. In our study, only the urinary recovery of xylose increased after fluid loading. These results suggest that fluid loading does not affect measures of intestinal permeability, assessed by the L/R ratio, but increases mediated carbohydrate transport in healthy volunteers, measured by xylose recovery.+ reabsorption and decreased pericapillary osmotic pressure with saline infusion, which might enhance the net movement of lactulose from the peritubular capillary into the lumen [In contrast with our findings, fluid loading has been shown to increase the L/R ratio, caused by increased urinary lactulose excretion, in both control and endotoxemic rats . Increashe lumen ,10. Systhe lumen ,11,12.In rats, Hallemeesch and colleagues  used douIt has been shown in volunteers that renal clearance of lactulose but not of rhamnose is dependent on the intravenously administered quantity of the respective sugar : urinaryXylose and 3-OMG are used to measure the mediated transport of carbohydrates and to assess the intestinal absorptive capacity. In animals, D-xylose is transported across membranes by a Na-dependent transport . HoweverIn critically ill patients, sugar absorption tests as a measure of gastrointestinal permeability have been challenged. In patients with multiple organ failure, urinary flow and creatinine clearance were the most important determinants of urinary sugar recovery . It was Our results suggest that fluid loading despite increasing mediated carbohydrate transport does not affect the L/R ratio after oral sugar administration in healthy volunteers. However, further studies on intestinal and renal sugar handling in the assessment of gastrointestinal permeability are needed in critically ill patients.• 	Fluid loading increases mediated intestinal carbohydrate transport in healthy volunteers.• 	Fluid loading does not affect intestinal permeability as assessed by the lactulose/rhamnose ratio in healthy volunteers.3-OMG = 3-O-methyl-D-glucose; L/R ratio = lactulose/rhamnose ratio.The author(s) declare that they have no competing interests.IP analysed data and wrote the final manuscript. JT designed the study and participated in the drafting of the manuscript. SJ recruited subjects into the study, managed them and participated in the drafting of the manuscript. All authors read and approved the final manuscript.


Recombinant human erythropoietin (rHuEpo) has recently become available for the treatment of chronic anaemia, including that associated with cancer. Carcinoma NT in CBA mice causes a progressive anaemia which can be overcome by daily injections of recombinant human erythropoietin (rHuEpo). This model was used to study the effect of haematocrit on tumour blood flow, growth rate and radiosensitivity, in mice with haematocrits ranging from approximately 38% (control) to 65% (20 U/day rHuEpo). Tumours showed a small but significant slowing in growth rate with higher haematocrit. In vitro studies showed rHuEpo had no direct effect on the growth of NT cells. Tumour blood flow was measured by two methods in each mouse (133Xe clearance and 86Rubidium uptake). Blood flow showed a tendency to decrease with increasing blood viscosity although this effect was not significant despite the large differences in haematocrit. Although tumour doubling time was prolonged despite the large differences in haematocrit. Although tumour doubling time was prolonged with increasing radiation dose, from 0 (sham irradiated) to 35 Gy, haematocrit was not found to influence the growth delay. This was attributed to adaptation of the tumour during the relatively slow change in the haematocrit. rHuEpo is being considered for clinical use in anaemic cancer patients. Our data suggest that this treatment will correct haematocrit with no effect on tumour radiosensitivity.


We have recently used two-dimensional DNA typing to detect genetic alterations in breast tumours. This method, which is based on size separation in neutral gels and sequence separation in denaturing gradient gels followed by hybridisation analysis with mini- and microsatellite core probes, allows the simultaneous analysis of hundreds of allelic fragments in a very short time. Here we demonstrate the potency of this method for total genome scanning of the tumour genome by analysing a small series of breast cancers. Comparison of tumour and normal DNA from ten breast cancer patients, using two-dimensional DNA typing with four core probes, revealed a considerable number of genomic alterations. In contrast, with Southern blot analysis only a few alterations were observed using the same probes. Most of the changes observed (74%) were deletions (absence of spots in the tumour) while 20% corresponded to amplifications (spots of higher intensity in the tumour) and 5% were new spots (gains). About 10% of the genomic changes detected appeared to occur in the tumours of more than one patient.


PCR-based microsatellite polymorphisms have proved their power in genetic linkage analysis and other identification methods, due to their high information content and even distribution over the chromosomes. In the present study we applied microsatellite polymorphisms to detect loss of heterozygosity in fresh (snap-frozen) and in archival ovarian tumour tissue. Clear allele losses were found in fresh and paraffin embedded tumour samples. Conventional Southern analysis of flanking markers on the same tumour DNA samples confirmed the observed losses detected by microsatellite polymorphisms. Titration experiments suggest that loss of heterozygosity remains detectable in tumour samples despite 60% contamination with normal DNA. This technique provides a fast and reproducible alternative to conventional Southern blotting in the detection of loss of heterozygosity, with the crucial additional advantages of minimal sample requirements, making archival material available for genetic investigation.


The effect of intraperitoneal (i.p.) injection of recombinant human interleukin-1 beta (rhIL-1 beta) was studied in three i.p. nude mouse xenograft models of human ovarian cancer . Intraperitoneal rhIL-1 beta administration led to a dose dependent replacement of peritoneal ascitic tumour with solid tumours attached to the peritoneum and intraabdominal viscera in two (HU and LA) out of the three xenograft models. In the third xenograft model (OS), low doses of rhIL-1 beta (10 ng day) promoted micrometastatic peritoneal implants of tumour, but higher doses of rhIL-1 beta (1 microgram day) had a marked antitumour effect. This was due to direct cytotoxicity for tumour cells and was not related to peritoneal neutrophil influx induced by rhIL-1 beta. Recombinant human TNF (rhTNF) also promoted tumour implantation in all three xenograft models, but its antitumour effects differed from rhIL-1 beta. TNF increased the survival of HU and LA bearing mice, but had no antitumour effect in the OS xenograft model. Analysis of peritoneal fluid and tumour xenografts showed that TNF induced murine IL-1 in the tumour bearing mice. The magnitude of IL-1 induction indicated that TNF induced IL-1 did not contribute significantly to its effects.


A study has been carried out in which a comparison was made between EL4 lymphoma (assumed to be an insulin-producing secreting tumour) and thymoma (an insulin-dependent tumour). Tumour development and incidence, 3H-thymidine incorporation and insulin content in tumours, the host's food intake, blood insulin, glucose and cholesterol were determined in non-diabetic and alloxan-diabetic mice. Whereas no significant differences were observed between the diabetic and non-diabetic EL4 tumour-bearing mice, the diabetic, thymoma tumour-bearing mice showed reduced tumour growth and lower tumour incidence as compared with their non-diabetic counterparts. Insulin administration to diabetic tumour bearing mice, enhanced 3H-thymidine incorporation in the thymoma tumour cells only, and the insulin content of the EL4 tumours was found to be higher than that of the thymoma tumours. Rapid diabetes remission was observed in the diabetic, EL4 tumour-bearing mice as compared with the thymoma tumour-bearing mice.


Monoclonal antibody (MAb) E48 and its F(ab')2 fragment, radiolabelled with 131I, were tested for tumour localisation and imaging in nude mice bearing a squamous cell carcinoma xenograft line derived from a head and neck carcinoma (HNX-HN) or from a vulva carcinoma (VX-A431). MAb IgG or F(ab')2 fragments were injected in parallel and at day 1, 2, 3 and 6 or 7, mice were either scanned with a gamma camera or dissected for determination of isotope biodistribution. In HNX-HN bearing mice, E48 IgG as well as F(ab')2 showed highly specific localisation in tumour tissue. The mean tumour uptake (n = 4) expressed as the percentage of the injected dose per gram of tumour tissue (percentage ID/g) of IgG was 11.9% at day 1 and increased to 14.6% at day 6 whereas percentage ID/g of F(ab')2 was 7.2% at day 1 and decreased during subsequent days. Tumour to blood ratios (T/B) at day 1 were 1.2 for IgG and 13.6 for F(ab')2 and reached a maximum at day 6 with values of 6.4 and 54.2 respectively. In VX-A431 bearing mice, only E48 F(ab')2 showed preferential localisation in tumour tissue. At day 1, Percentage ID/g of IgG was 3.7 and T/B was 0.3, while percentage ID/g of F(ab')2 was 2.4 and T/B was 3.2. Percentage ID/g decreased after day 1 while T/B increased. In these experiments no preferential localisation of either isotype matched 125I-labelled control IgG or F(ab')2 was observed. In F(ab')2 injected HNX-HN bearing mice as well as VX-A431 bearing mice, tumours could be visualised at day 1 and 2 without any appreciable background activity. With MAb IgG this was also possible in HNX-HN bearing mice (but not in VX-A431 bearing mice) but only at day 3 and 6. These findings suggest that the superior tumour to non-tumour ratios render the E48 F(ab')2 fragment more qualified for specific targeting of radioisotopes to tumour xenografts in this experimental setting.


A new bisphosphonate, disodium dihydrogen (cycloheptylamino) methylene bisphosphonate monohydrate (YM175), was compared with 3-amino-1-hydroxypropylidene-1, 1-bisphosphonate (AHPrBP) and 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) in terms of its effect on tumour induced osteolysis using a bladder tumour in mice (MBT-2). The method consisted of inoculating tumour cells subcutaneously (SC) over the calvaria in mice, resulting in a local tumour causing fragmentation of the bone. The compounds were active not only when administered preventively before establishment of bone resorption, but also in an inhibitory fashion once the variables were already under the influence of the tumour. This osteolysis was evaluated by measuring the increased area of bone resorption in reduced opacity to radiograph and histology. The results showed the following sequence of potency: YM175 > AHPrBP = HEBP. This inhibition was obtained with no apparent effect on the growth of the MBT-2 tumour. YM175 appears to be an interesting new bisphosphonate with possible clinical application.


EMT6 multicellular spheroids were introduced into the peritoneal cavities of mice and allowed to become vascularised, resulting in solid spherical tumours. The necrotic cores of the initially avascular spheroids were replaced by vascularised tumour tissue but the outer zones of the spheroids failed to become vascularised. The presence of both vascular and avascular components in each spheroid allowed the role of the vasculature in the antitumour action of flavone acetic acid (FAA) to be determined. Eighteen hours after treatment with FAA 0.8 mmol kg-1, the vascularised core became necrotic and haemorrhagic, while the outer avascular zone remained viable. Tumour cells which were infiltrating superficial sub-mesothelial fat did not become necrotic despite the presence of numerous thrombi in associated vessels. Injection of two fluorescent vascular markers, the first (Hoechst 33342) together with FAA, and the second (10-nonyl acridine orange) 4 h later, demonstrated that there is a marked loss of blood flow in the spheroids. These results provide further evidence that FAA kills blood vessel-dependent tumour cells by interrupting the tumour blood supply.


N-l-leucyl-doxorubicin and vinblastine-isoleucinate can be considered as relatively non-toxic prodrugs from doxorubicin and vinblastine, respectively. A comparative analysis was carried out of the anti-tumour activity of the four compounds as well as vintriptol in four human ovarian cancer xenografts different in histology, growth rate and chemosensitivity. Injections were given i.v. weekly twice into mice bearing well-established s.c. tumours. At equitoxic doses, the amount of drug administered for N-l-leucyl-doxorubicin and vinblastine-isoleucinate was respectively 3-fold and 2-fold higher than the doses of the parent compound. N-l-leucyl-doxorubicin induced a growth inhibition > 50% in three out of four human ovarian cancer lines. The anti-tumour effects obtained were significantly better (P < 0.01) than in the case of doxorubicin. Vinblastine-isoleucinate studied in two of these lines could induce a growth inhibition of > 50%. This prodrug appeared slightly less effective than vinblastine. Insignificant growth inhibition (< 50%) was obtained by vintriptol.


The growth kinetics of established human colorectal tumour cell lines  and human diploid fibroblasts (Flow 2002) were studied in conventional culture and in microcapsules formed from alginate-poly(L-lysine)-alginate membranes. The tumour lines grew rapidly in microcapsules but, in the case of the substrate-adherent lines HT29 and HT115, only after a prolonged lag phase. This phase was reduced by serial passage in microcapsules. The anchorage-independent line COLO 320DM showed no lengthening in lag phase. Microencapsulated fibroblasts underwent negligible growth but remained viable. Some evidence for functional differentiation  of the tumour line HT115 within the microcapsules was observed. We conclude that the use of microcapsules provides an alternative system with some advantages for the study of human cancer and its metastases in vitro.


Commonly accepted outcomes of varicella-zoster virus (VZV) infections include chickenpox (primary) and shingles (recurrence or latency), as well lifetime immunity against chickenpox. We report the case of a registered nurse who worked in a neurologic surgery ward in a general hospital in Taipei, Taiwan. While working there for approximately 1 year, she developed recurrent chickenpox after caring for a paraparesis patient, who had herpes zoster during hospitalization in August 2002. The varicella incubation period was 10 days, which matched the range (10–21 days). Recently negative specific serum IgM and positive specific serum IgG indicated a past VZV infection. The nurse did not get herpes zoster from the second episode of varicella on 9 August 2002 to 4 April 2005 and is now convalescing. We conclude that occupational VZV hazards exist in the health care environment and suggest testing for VZV antibody and a VZV vaccination program for susceptible health care workers. Varicella (chickenpox), a common contagious disease of childhood, is caused by the varicella zoster virus (VZV) . VZV is International Classification of Diseases, Revision 9 (ICD-9) code 052.9; A 25-year-old nurse, who had childhood chickenpox, was diagnosed with varicella without mention of complication , but the VZV IgG was positive , with high sensitivity (93.9% for IgM and 100% for IgG) and high specificity (100% for both antibodies) according to the manufacturer’s instructions.Negative specific serum IgM and positive specific serum IgG indicated a past VZV infection. Because varicella and herpes zoster both resulted from the same antigen, the VZV IgG-positive reaction was excluded from the herpes zoster. It was also excluded because the nurse did not get herpes zoster from the second episode of varicella on 9 August 2002 to 4 April 2005. The nurse is now convalescing.The nurse was diagnosed with an acute chickenpox infection by a dermatologic physician at the teaching general hospital on the basis of her clinical symptoms and signs, including a high fever (39.5°C), malaise, and headache prior to the rash appearing on her face, neck, and trunk. The duration of the high fever and rash were 5 and 14 days, respectively. Fever and malaise may appear 1–2 days before rash onset in chickenpox primary infections in adults, whereas rash is usually the first sign of the disease in children . AccordiThe air sampling study was performed on 9 July 2002 in the neurologic surgery ward, and the paraparesis patient was admitted to the hospital on 13 July 2002 and transferred to the neurologic surgery ward on 18 July 2002. The patient’s rash was first reported on 3 August 2002. It is not surprising that the air sampling result was negative. In addition, it is unlikely that the nurse was infected from the air sampling equipment because the assembled cassettes and sampling pump with tubing were put in a laminar flow unit with ultraviolet light for 8 hr before sampling, and sampled cassettes were replaced with a new assembled cassette every day.To investigate this uncommon case, we considered four parameters: the agent (VZV), infectious routes, the host (the nurse), and the hospital environment .Previous reports have mentioned the same etiology of varicella and herpes zoster . In 1925Varicella has been reported as the disease most commonly confused with smallpox ; howeverNew variants of VZV have been reported recently, including VZV-MSP, isolated in St. Paul–Minneapolis, MN , 2000, aIt has been documented that cases of herpes zoster are the infectious sources of chickenpox in susceptible persons . VZV is Previous PCR research showed that VZV is transmissible for 24–48 hr before rash onset, which is consistent with epidemiologic evidence , or 4–5 The chickenpox incubation period is from 14 to 16 days after exposure , with a In addition to a lack of protection  while caring for the patient, stress may be another important host factor contributing to recurrence. The nurse reported that before she resigned from the hospital, she felt stress both from the nursing practice and the logistics of her resignation, as well as anxiety about her impending study in graduate school. In fact, during our study of occupational VZV hazards to healthcare workers in early July 2002, the nurse reported performance anxiety about graduate school and stress from multiple clinical duties and logistic matters concerning her resignation, such as the fact that she was on duty on the date of graduate school registration, 5 August 2002. According to the nurse’s own report after the recurrence, we believe that stress played an important role in this recurrent varicella.Documented environmental VZV DNA included sampling from the air of the active patient’s room , or the The nurse was a young and unmarried female, with only one year of work history. Her disease appears to be recurrent varicella, which is very uncommon, and it appears to be occupationally related. Documented VZV infection may cause significant morbidity or mortality, affecting the mother, the fetus, or the newborn , as wellDocumented methods of VZV antibody detection include complement fixation, IFA, fluorescent antibody to membrane antigen (FAMA), neutralization, indirect hemagglutination, immune adherence hemagglutination, radioimmunoassay, latex agglutination, and enzyme-linked immunosorbent assay . AlthougOccupational VZV hazards exist in the health care environment. The traditional concept of VZV lifetime immunity after the primary varicella infection may not be appropriate in a health care setting. We suggest checking serologic titers for VZV antibody, followed by a VZV vaccination for nonimmune health care workers.


The present investigation shows the effect of vitamin A deficiency on the initiation and postinitiation phases of benzo(a)pyrene-induced lung carcinogenesis in male Wistar rats. Lung tumours were induced by giving three intratracheal instillations, one week apart, of 10 mg benzo(a)pyrene per instillation. Maximum tumour incidence (100%) and tumour burden per rat was found in rats which were kept on vitamin A deficient diet for 4 weeks prior to the first administration and 8 weeks after the last administration of benzo(a)pyrene. Rats in which vitamin A deficiency was terminated after the last administration of the carcinogen had 83% tumour incidence, whereas corresponding control pairfed animals had 39% incidence of tumours. These data represent the values obtained 32 weeks after the last administered dose of the carcinogen and indicate the role of vitamin A, both in the initiation as well as in the postinitiation phases of lung carcinogenesis.


The candidate tumour-suppressor gene, LUCA-15/RBM5/H37, maps to the lung cancer tumour-suppressor locus 3p21.3. The LUCA-15 gene locus encodes at least four alternatively spliced transcripts that have been shown to function as regulators of apoptosis, a fact which may have major significance in tumour regulation. This review highlights recent evidence that further implicates the LUCA-15 locus in the control of apoptosis and cell proliferation, and focuses on the observations that confirm the tumour-suppressor activity of this gene.


We studied the mouse NC tumour, a subcutaneously transplanted adenocarcinoma originally of mammary origin. Measurements per g tissue were made of 17 fatty acids (FAs), the combined amounts of n-3, n-6, saturated, unsaturated, and total FAs, and of various FA ratios in the tumour, mammary tissue, spleen, liver and plasma. Compared with mammary tissue from normal mice, tumours of vehicle-treated controls had less of seven of the FAs and more of two FAs. Mice bearing the NC tumour often had changed  amounts of FAs in the 'normal' spleen, liver and plasma, but not in mammary tissue. Treatment with methotrexate (MTX) was studied alone and with indomethacin which can potentiate MTX cytotoxicity. Indomethacin 1.25 mg kg-1 (INDO) increased the amounts of 3/17 tumours FAs and the unsaturated FAs, but reduced 9/17 FAs, the saturated and the unsaturated FAs in 'normal' mammary tissue, and usually had no effect on the FAs of other tissues. MTX 2 or 4 mg kg-1 (MTX 2 or 4 mg) +/- INDO in general partly restored (increased) the amounts of tumour FAs, and reduced the saturated/unsaturated FA ratio. In the 'normal' spleen and plasma also, but not in the liver, MTX 2 mg generally somewhat restored the FA composition. However, as in the liver, the spleen 20:4 and 22:6 (which form prostaglandins and lipid peroxides) did not increase in the presence of INDO. With MTX 4 mg, some of the plasma and liver FAs decreased, in contrast to the tumour. There was generally no evidence of MTX potentiation by INDO. These results are discussed in relation to carcinogenesis, cachexia, and the response to treatment.


WHT/Ht mice transplanted s.c. with NC carcinoma were treated with 16,16-dimethyl prostaglandin E2 methyl ester (di-me-PGE2) and/or indomethacin. Each primary tumour was excised under anaesthesia 3 weeks after transplantation, weighed and extracted for prostaglandins. Mouse survival time and tumour recurrence were measured. Di-me-PGE2 10 micrograms, injected at the tumour site on alternate days from day 1 to 19, indomethacin 2.5 mg kg-1 daily by mouth, or both drugs together resulted in lighter tumours  compared with vehicle-treated controls. Indomethacin reduced the tumour prostaglandin yield, but the biological activity in extracts of tumours from mice given di-me-PGE2 was high. The median survival time was longer in mice receiving indomethacin alone (61 days from tumour transplantation compared with 50 days in controls P less than 0.02). Di-me-PGE2 alone had little or no effect on survival (median 48 days) but counteracted the increase with indomethacin . There were no obvious effects of the treatments on tumour recurrence at the excision site, but there was a higher incidence of involved lymph nodes in mice given di-me-PGE2.


FCE 23762 is a new doxorubicin derivative obtained by appending a methoxymorpholinyl group at position 3' of the sugar moiety. The compound is greater than 80 times more potent than doxorubicin, it is highly lipophilic, and presents equivalent anti-tumour activity when administered by i.p., i.v. or oral route. The pattern of anti-tumour activity of FCE 23762 differs from that of doxorubicin in maintaining anti-tumour activity against two P388 murine leukaemia sublines resistant to doxorubicin and, although at borderline levels of efficacy, against LoVo human colon adenocarcinoma resistant to doxorubicin. FCE 23762 exhibits remarkable efficacy against MX-1 human mammary carcinoma, with most treated mice being cured both after i.v. and oral treatment. Anti-tumour activity was also observed against L1210 murine leukaemia and two sublines resistant to cis-platinum and melphalan, M5076 murine reticulosarcoma, MTV murine mammary carcinoma and N592 human small cell lung cancer.


Recent circumstantial evidence has implicated Insulin-like growth factor II in the genesis of several tumour types, notably developmental tumours . This type of tumour, thought to originate during the defective differentiation of organ precursors , often expresses greatly elevated levels of mRNA for IGF-II, a known mitogen for these cells and abundantly expressed in their presumed normal counterparts . It has been proposed that continued, inappropriate expression of this gene drives tumour growth by an autocrine mechanism. In order to examine the potential role of IGF-II in the growth of tumour cells an IGF-II cDNA was introduced into a retroviral expression vector, and used to infect a cloned fibroblast cell line. Expression of IGF-II conferred a degree of serum independence of growth in cell culture, however, when cells were injected into nude mice as subcutaneous grafts, clones expressing IGF-II from the retrovirus were found to have a greatly increased (five fold) latency of sarcoma formation. After a prolonged lag all cell lines eventually gave rise to tumours in which the introduced IGF-II genes had either been lost or inactivated, suggesting that in this system IGF-II acts as a tumour suppressor gene.


Natural killer (NK) cells are involved in the antitumoral immunologic mechanism. These cells act through the release of cytotoxic molecules defined as NK cytotoxic factors (NKCF). Inhibitory factors of NK and NKCF mediated lysis have been described in in vitro assays. This study evaluates the induction of resistance to NKCF cytotoxicity by sera from 27 patients with colon, breast and lung cancer. Addition of these sera to the cytolytic assay where K562 cells and concentrated NKCF were used, induced resistance to NKCF mediated cytotoxicity in 21 cases (77%). The sera from the group with metastasis blocked NKCF lysis more markedly than the group with local tumours. However, no differences were observed when the groups with colon, breast and lung cancers were compared. This blocking effect was not found to be related to gamma interferon (IFN) levels. In a previous study, we described a tumour factor (NK-RIF) produced by human cell lines derived from metastatic adenocarcinomas. This factor blocked lysis of tumour target cells by NK cells. Consequently, it is proposed that the release of similar tumour factors with a capacity to induce resistance to NKCF may be involved in tumour growth and metastatic spreading in in vivo.


In an attempt to develop an assay to predict patient tumour response to cyclophosphamide (CP), the feasibility of using a glutathione-targeted assay to assess the in vitro chemosensitivity of tumour cells to 4-hydroperoxycyclophosphamide (4-OOH-CP), an activated congener of CP, was evaluated. A panel of 19 human and three murine tumour cell lines was used. These consisted of three main categories of tumour types, viz. ovarian, lung and squamous cell carcinoma. The major finding was that the occurrence of a significant reduction of tumour cell reproductive capacity was always accompanied by substantial depletion of cellular glutathione (GSH) content, and vice versa. Plots of % GSH depletion versus clonogenic cell survival demonstrated highly significant correlation . It was determined that for in vitro tumour cell lines, a GSH depletion to 40% of initial content may serve as a cut-off criterion for chemosensitivity to 4-OOH-CP. This degree of GSH depletion is indicative of clonogenic cell survival of approximately 1% (95% confidence limits = 3 x 10(-5)-1.6 x 10(-2)). The relationship between steady state GSH content and intrinsic sensitivity to 4-OOH-CP was also evaluated. The GSH concentration of the tumour cell lines ranged from 1.3-21.2 x 10(-18) moles microns-3; chemosensitivity to 4-OOH-CP, in terms of IC99, was in the range of 5.0-87.1 microM. A good correlation was observed between these two parameters . These results suggest that GSH plays an important role in determining the therapeutic efficacy of 4-OOH-CP in the treatment of cancer. It is uncertain, however, whether a high tumour steady state GSH content in itself is sufficient to cause therapeutic failure in patients.


The suggestion that the ketonaemic response to fasting may be altered in the tumour-bearing state was investigated by studying the metabolism of fasted C57/BL6j mice bearing transplanted B16 melanomas. Ketone body (D-3-hydroxybutyrate and acetoacetate) concentrations in the blood of the tumour-bearing mice were significantly increased after a 24 h fast compared to control mice with identical dietary histories. Hepatic glycogen levels were lower at the start of the fasting period in the tumour-bearing mice as were the fat stores. The loss of adipose tissue during the fasting period was greater in the tumour-bearing mice. After 48 h of fasting, the ketonaemia was significantly lower in the tumour-bearing mice compared to the appropriate controls. Two distinct metabolic states are indicated in these fasted tumour-bearing mice, one characterised by accelerated ketonaemia, and a later, near terminal stage, where fat deposits are markedly depleted and ketonaemia is decreased.


The glutathione S-transferases (GSTs) have been implicated in carcinogenesis and tumour drug-therapy resistance. In this study GST pi was the predominant isoenzyme in the fetal human kidney. It was present in differentiated epithelial structures but never in the primitive mesenchyme. By contrast most cases of Wilms' tumours showed GST pi in both epithelial structures and undifferentiated blastema. The level of expression, as assessed by immunostaining, was no more than moderate, and was generally higher in differentiated elements. In only one case was GST alpha found in Wilms' tumour. This study had demonstrated a difference between fetal kidney and Wilms' tumour blastema in terms of GST expression.


The importance of host immune status on the spontaneous metastasis of cloned cell lines of the 13762NF rat mammary adenocarcinoma was examined. Cell lines MTLn3 , MTF7 and MTLn2  and MTC  were subjected to a series of in vivo assays designed to assess how manipulation of the immune system in the syngeneic F344 host would affect the ability of these cells to metastasise. Treatment of tumour bearing rats with the immunosuppressive agents cyclosporin A or cyclophosphamide had little influence on metastasis in this system. Growth of tumours in congenitally athymic nude rats resulted in reduction of observed metastases. In addition, humoral immune response was not detectable during a 23-day period of tumour growth in F344 rats. Excision of the tumour growing in situ reduced the number of metastases when the tumours were resected early (less than 10 days), but at later times tumour resection did not influence the incidence of metastasis. The importance of initial lymphatic rather than haematogenous routes of dissemination was confirmed in experiments where the draining inguinal and axillary lymph nodes were removed at different times either before, or after, subcutaneous mammary fat pad injection of metastatic tumour cells.


To investigate the influence of hormones on the process of cellular differentiation the growth and differentiation of a transplantable tumour, induced by inoculation of pluripotent mouse embryonal carcinoma (EC) cells have been studied in athymic nude mice and, normal and hypopituitary Snell dwarf mice. All athymic nude mice developed tumours independent of the numbers of cells inoculated. In contrast, the tumour percentage in normal Snell mice was lower, showing a dose-dependent increase of takes. In dwarfs tumour percentage was comparable with that observed in normal Snell mice. The morphological differentiation of teratocarcinomas grown in athymic nude mice, normal and dwarfed Snell mice shows derivatives of all three germ layers next to undifferentiated embryonal carcinoma cells. This suggests that the pituitary hormonal deficiencies of the dwarfs  did not influence the tumour induction nor the development of the different tissues present in this type of tumour.


Previous studies showed that murine polymorphonuclear leukocytes (PMNs) lyze tumour cells in the presence of wheat germ agglutinin or actinomycin D. This paper reports studies on whether a soluble factor participates in PMN-mediated cytolysis dependent on lectin or a chemotherapeutic drug. Tumour lysis was observed with supernatants from PMN cocultured with wheat germ agglutinin or actinomycin D. The supernatant from cultures of PMNs alone was not cytotoxic, but addition of these agents to the supernatant induced tumour lysis. PMNs released a soluble factor spontaneously into the medium and cytolysis was induced by a combination of this factor and wheat germ agglutinin or actinomycin D. This factor was not an oxygen metabolite, but a protein with a molecular weight of approximately 100 K daltons. These results suggest that a soluble factor(s) from PMNs participates in tumour killing in cooperation with appropriate reagents.


The prognostic significance of argyrophilic nucleolar organiser regions (AgNORs) has been evaluated in biopsy specimens from 61 primary squamous and undifferentiated carcinomas of the pharynx prior to therapy. The univariate Kaplan-Meyer survival analysis showed a significant correlation between 3- and 5-year survival rates and the mean AgNOR number per tumour cell (P less than 0.001). No significant correlation was found between prognosis and patients age and sex, tumour location, clinical stage, histologic grade, extent of lymphocytic infiltration, HMFG-2 positivity of tumour cells and UCHL1, LN2, MB2 positivity of infiltrating lymphocytes. There was no significant association between AgNOR counts and tumour histologic grade or clinical stage. Multivariate survival analysis showed that only two variables were significantly correlated with prognosis: AgNOR counts (P less than 0.001) and the extent of lymphocytic infiltration (P less than 0.027). Our results indicate the prognostic value of AgNOR counts and suggest the use of this method as a significant parameter in the pretherapeutic assessment of the aggressiveness of pharyngeal carcinomas.


An oestrogen-sensitive rat mammary tumour (OES HR1) has been grown in normal female rats and in female and male rats supplemented with oestrone. In some rats, after the tumour was established, both exogenous and endogenous sources of oestrogen were removed--a treatment which inhibited further growth of the tumour. The proliferative characteristics of the tumours were measured by injecting the rats with deoxybromouridine (BrdU) 4 h before removing the tumour. Extracted nuclei were reacted with anti-BrdU and the labelling index and DNA content measured by flow cytometry. A correlation between the number of (diploid) host cells present and the number of (aneuploid) tumour cells in S-phase of the cell cycle was observed. This result suggests that there are paracrine interactions between tumour and host cells. We also observed that, on oestrogen ablation, the labelling index was significantly reduced while the percentage of cells in S-phase changed far less. The demonstration that there are cells in S-phase which are not proliferating highlights a possible problem with the measurement of proliferation in human tumours from a DNA histogram.