Quantitative measurement of gene expression of the genes causing hepatocellular carcinoma

Hepatocellular carcinoma (HCC), the major type of primary liver cancer, is one of the most common malignancies worldwide.This malignant tumor develops on a back ground of chronic hepatitis or cirrhosis. Effective therapy to this cancer is currently lacking, creating an urgent need for new therapeutic strategies for HCC.The aim of this study was therefore to investigate the effect of vitamin D2 and free fatty acids (FFA) on stress ligands (MICA and MICB) and pro-fibrotic activity (?-SMA COL1? and TGF-ß) in hepatic stellat cells (LX-2 cells and human primary HSC) and hepatocellular carsinoma cells (HepG2, Huh7). LX-2 cells were treated with VD2 (10-6 M) and 1 mM FFA for 24 hours, primary human HSC were treated with VD2 (10-6 M) and 0,25 mM FFA for 24 hours, HepG2 and Huh7 cells were treated with VD2 (10-6 M) and 0,5 mM FFA for 24,48 and 72 hours and and expression of the genes were assessed by quantitative real time-PCR. In summary this study shows that compared to vehicle controls, treatment of cells with 0.5 mM and 1 mM FFA induced ? -SMA and TGF-ß expression in LX-2 cells. VD2 treatment significantly downregulated the FFA-induced expression of TGF-ß and ?-SMA in LX-2 cells. COL1? expression was reduced after addition of FFA. Moreover, VD2 treatment significantly downregulate expression of MICA in isolated primary HSC and MICA/B expression in primary HSC was not affected by FFA treatment. Also, in HepG2 cells a significant decrease of MICA and MICB mRNA by VD2 was detectable. Following VD2 treatment for 72 hours of Huh 7 cells significantly downregulate expression of MICB. These results indicate that vitamin D2 may reduce inflammatory and profibrogenic activity of stellate cells in vitro.Key Words: Hepatocellular carcinoma, Hepatic stellate cells, MICA/B, Vitamin D2