Effects of JS K, a novel anti cancer nitric oxide prodrug, on gene expression in human hepatoma Hep3B cells.
JS K is a novel anticancer nitric oxide (NO) prodrug effective against a variety of cancer cells, including the inhibition of AM 1 hepatoma cell growth in rats. To further evaluate anticancer effects of JS K, human hepatoma Hep3B cells were treated with JS K and the compound control JS 43 126 at various concentrations (0 100mM) for 24h, and cytotoxicity was determined by the MTS assay. The compound control JS 43 126 was not cytotoxic to Hep3B cells at concentrations up to 100mM, while the LC50 for JS K was about 10mM. To examine the molecular mechanisms of antitumor effects of JS K, Hep3B cells were treated with 1 10mM of JS K for 24h, and then subjected to gene expression analysis via real time RT PCR and protein immunostain via confocal images. JS K is a GST a targeting NO prodrug, and decreased immunostaining for GST a was associated with JS K treatment. JS K activated apoptosis pathways in Hep3B cells, including induction of caspase 3, caspase 9, Bax, TNF a, and IL 1b, and immunostaining for caspase 3 was intensified. The expressions of thrombospondin 1 (TSP 1) and the tissue inhibitors of metalloproteinase 1 (TIMP 1) were increased by JS K at both transcript and protein levels. JS K treatment also increased the expression of differentiation related genes CD14 and CD11b, and depressed the expression of c myc in Hep3B cells. Thus, multiple molecular events appear to be associated with anticancer effects of JS K in human hepatoma Hep3B cells, including activation of genes related to apoptosis and induction of genes involved in antiangiogenesis and tumor cell migration.
Biomedicine pharmacotherapy Biomedecine pharmacotherapie