Still, the underlying mechanisms promoting HCC development and progression in NAFLD are only incompletely understood. The aim CHIR-99021 purchase of this study was to analyze the influence of high free fatty acid levels and hepatic steatosis on HCC. Methods and Results: We applied a syngeneic, orthotopic HCC model, in which we implanted murine Hepa129 HCC cells into the liver of C3H/He mice, which (i) had been fed with a high-fat diet (HFD) causing significant hepatic steatosis and (ii) control mice, fed with standard chow and normal liver histology. HFD feeding was continued until mice were sacrificed 14 days after tumor cell implantation. Tumors
formed in steatotic livers were significantly larger, showed less apoptosis and revealed an invasive growth, while tumors in the control group showed no infiltration in the surrounding liver parenchyma. Tumors grown in steatotic environment also revealed higher expression
of matrix metalloproteases (MMPs) and the anti-apoptotic gene survivin, and interestingly, also had a higher triglyceride content. HFD-fed mice had higher circulating free fatty acid (FFA) levels than control mice, and in vitro, addition of FFA to the cell culture medium caused a dose dependant induction of cellular lipid accumulation in Hepa129 cells. Steatotic HCC cells showed migratory activity in Boyden Chamber assays and higher resistance against apoptosis in FACS analysis. Conclusions: Hepatic MK-2206 datasheet steatosis does not only induce the growth but also the invasiveness of HCC cells, indicating the importance of a steatotic environment for HCC progression. In addition, systemic metabolic changes associated with the metabolic syndrome such as dyslipidemia seem to directly affect HCC cells offering a further option for HCC prevention and treatment. Disclosures: The following people have nothing to disclose: Andreas Koch, Claus Hellerbrand Background: Glutathione peroxidase 4 (GP×4) is a selenium containing antioxidative enzyme with a unique function to eliminate lipid hydroperoxides. The involvement of GPx4 in carcinogenesis has been 上海皓元医药股份有限公司 shown for colon cancer, but the role
of GP×4 in hepatocellular carcinoma (HCC) remains to be investigated. Methods: Expression plasmids with the porcine GPx4 gene under control of the CMV promoter were transfected into human Huh7 and HCC-3 hepatocarcinoma cell lines. The transfection efficiency was evaluated by real-time PCR, by western blotting, and by activity measurements. The effect of GP×4 on tumour growth and vascularisation was assessed by xenotrans-plantation into NSG recipient mice. After immunohistochemical staining, the expression of GPx4, cell proliferation and vessel formation were determined by histomorphometric analysis of paraffin embedded tumour tissues. The expression of angiogen-esis-related genes was measured by real-time RT-PCR.