acute myeloid leukemia ( aml ) is a malignant clonal disorder of immature myeloid hematopoietic cells that is the most common type of acute leukemia among adults . reported mean age of aml patients is 60 . some factors such as viruses , radiation , chemotherapy , benzene exposure and smoking increase risk of aml ( 1 - 4 ) . also , agents such as age , molecular and genetic changes such as flt3 , npm1 , mll disorders and hematologic indices are important prognostic factors in aml process ( 5 - 11 ) . in addition to mentioned genes , brain and acute leukemia - cytoplasmic ( baalc ) gene is an important factor in treatment and prognosis determination ( 12 - 16 ) . overexpression of baalc gene is reported in aml subjects with trisomy 8 abnormalities in gene expression studies ( 12 ) . it is a new marker in hematopoietic progenitor cells , which its effects are indeterminate on cellular mechanism ( 12,13,17 - 19 ) . different studies emphasis on gene expression level impression on different physiologic and pathologic circumstances prognosis . generally , baalc expression is a suitable prognostic marker which can be used to assign survival rate in normal cytogenetic aml ( cn - aml ) individuals ( 12,14 ) . no study was found in aml patients in mentioned field in north east of iran , thus , the aim of this study was to define amount of gene expression and frequency of up and down regulation of baalc gene , and to determine the survival rate of affected patients . in this case - control study , 94 individuals referred to molecular pathology research center of ghaem hospital in mashhad city during 2012 - 2015 were recruited in 2016 . from all , 47 cases were normal cytogenetic aml and others were healthy individuals that were studied as control group . all patients were categorized based on french - american - british ( fab ) criteria . extracted information included peripheral blood and bone marrow blast count , red and white blood cells and platelets , mean cell volume ( mcv ) , mean cell hemoglobin ( mch ) , mean cell hemoglobin concentration ( mchc ) and clinical signs including splenomegaly , hepatomegaly , and lymphadenopathy and bleeding protests . it is necessary to mention that this research was approved by ethics committee of mashhad university of medical sciences , code 839 . peripheral blood mononuclear cell were separated by ficoll gradient centrifugation method and sigma aldrich kit ( sigma aldrich company , saint louis , mo , usa ) . in the following rna extraction was done by tri - pure solution ( tri - pure kit , cat no . revertaid h minus first strand cdna synthesis kit ( fermentase company , thermo scientific 100 reactions ) was applied for cdna synthesis . gpi gene control primers were evaluated for cdna qualification confirmation , by pcr method ; the results were checked on agarose gel . qualification of synthesized cdna was evaluated by nano drop instrument ( thermo scientific 2000 , made in finland ) in 260 nm wavelength. 
 
 all samples concentration was > 450ng/ l . studied genes sequences were obtained from baldus .et al study ( 13 ) . to determine primers specificity and evaluation of sequence homology blast software were applied which is available in ncbi site . forward primers for gpi gene were 5'- tcttcgatgccaacaaggac-3 ' and 5'-cttttgcaggcattctcttagca-3 ' for baalc gene . the reverse primers was 5-gcatcacgtcctccgtcac-3 ' for gpi and 5'-cttttgcaggcattct cttagca-3 ' for baalc gene . the used probe sequences were 5-ttcagcttgac cctcaacaccaac-3 ' for gpi and 5'-ctcttttagcctctgtggtctgaaggccat-3 ' for baalc. 
 
 real time pcrs were applied in thermo cycler abi ( made in usa ) . all samples were tested in duplicate . rt - pcr reaction master mix ( 20l for each reaction ) contained 10l taq man master mixes , 0.5l rox stain , 1l of each forward and reverse primers ( 10 pm ) , 1l taq man probe ( 5 pm ) , 2l cdna , 4.5l double distilled sterile water .finally the following temperature patterns were planned in termocycler ; 50c for 2 minutes , 95c for 15second , 95c for 15 seconds , and 60c for 1 minute . the second , third and fourth steps were repeated for 40 times . melting curve analysis was done for products specificity confirmation. 
 
  chi - square , t - test , kruskal - wallis and mann- whitney tests were applied for analysis based on data distribution ( checked using kolmogorov - smirnov test ) , through spss v.21 . there were 26 ( 55.3% ) male and 21 ( 44.7% ) female in patient group . patients were categorized to child and adult ; the first one was 15 and consisted of 13 ( 27.5% ) and the second was > 15 which consisted of 34 ( 72.5% ) approached subjects . most of patients were in m1 ( 29.78% ) and m2 ( 27.65% ) subtypes based on fab categorization . mean of peripheral blood and bone marrow blasts were 39.0% and 66.2% in aml cases , respectively . mean of hb , hct and rbc indices were significantly different between the two groups ( p=0.03 ) , which is justifiable with occurred pancytopenia . in comparison of baalc gene expression there was no any significant relation between amount of gene expression and clinical and hematological parameters ( p=0.02 ) . among hematologic indices , rbc count had significant invert relation with amount of baalc gene expression ; patients ' age and sex were related to baalc expression too ( p>0.05 ) . gene evaluation was associated with up regulation in 47% of cases and down regulation in 53% of patients . survival rate was assessed during 36 months in 47 aml patients and it was 26 months in down regulation subjects and 33 months in up regulation individuals . there was significant relation between up regulation of baalc and survivability reduction ( p=0.04 ) . in the recent decades , genomic profile has been used to identify prognostic factors of leukemia . erg , baalc , mn1 and wt1 genes are considered as categorization indices based on gene expression patterns ; they can be used to distinguish therapeutic intervention and disease process in aml patients ( 13,20 ) . although baalc overexpression mechanism has not been recognized yet , but it is reported in myeloid leukomogenesis . this up regulation prevents myeloid blast differentiation and is associated with poor prognosis in aml and acute lymphoblastic leukemia ( all ) individuals . different research revealed that baalc up regulation has been observed in glioblastoma , melanoma and gastrointestinal cancers ; they suggested an oncogenic role for baalc ( 21 ) . this gene expression is known as a prognostic and survival rate marker in normal cytogenetic aml cases ( 12 ) . baalc up regulation was seen in 47.00% of aml group members of this study ; damiani .et al research on 175 aml patients by rt - pcr method revealed up regulation in 50% of studied cases , and recommended it as a suitable marker for minimal residual disease ( mrd ) assignment ( 15 ) . high baalc expression was reported in 48.51% of 125 cn - aml individuals of yoon 's study in korea too ( 22 ) . baldus study on baalc gene expression and flt3 mutation in normal cytogenetic aml patients under 60 years found no significant differences between age , sex and bone marrow blast count and amount of expressed baalc gene , before treatment ; he mentioned poor prognosis of baalc up regulation too ; while cited gene overexpression and peripheral blood blasts increment were seen in m1 and m0 subtypes . in current study , parts of our findings were supported by baldus ( 13 ) and weber 's ( 12 ) studies which expressed that there were no any significant relationships between baalc expression , gender and hematological parameters ( wbc , hb , bone marrow and peripheral blood blasts count ) . yahya rs et al in their study about prognostic role of baalc gene in aml patients in egypt in 2013 represented up regulation in 48.9% of studied cases ; while they did not report any relations between baalc up regulation and clinical parameters . they reported baalc up regulation in association with patient 's survival rate reduction and indicated to its prognostic role ( 16 ) . we calculated amount of cited gene expression in each morphologic group ( based on fab ) , the most up regulated cases were m1 and m2 ; as mentioned before it is reasonable according to blast count in these groups . these findings are similar to baldus .et al reported result about normal cytogenetic aml patients under 60 ; as it mentioned before they observed overexpression of baalc along with peripheral blood blast increment in m1 and m0 and down regulation in m5b subtypes . their findings support our results about poor prognosis of baalc over expression too ( 13 ) . liu et al stated that baalc overexpression is seen in undifferentiated cells ; they found the most up regulation in m0 , m1 and m2 categories too , same as our findings ( 23 ) . current study emphasized that baalc up regulation is increased in aml patients , and it is more common in m1 and m2 subtypes .