The Effects Of Hepatic Stellate Cells With Overexpression G Protein-coupled Receptors Mas On Hepatocellular Carcinoma Cells

Objective To clarify the effect of hepatic stellate cells with overexpression G protein-coupled receptors Mas on hepatocellular carcinoma cells and reveal the effect of Mas receptor on hepatocellular carcinoma cells in tumor micro-environment,and provide a theoretical basis for future studying the exact mechanism of Mas receptor in hepatocellular carcinoma and possibility of Mas as the therapeutic targets of hepatocellular carcinoma.Methods The expression of Mas receptor and ?-SMA in human hepatocellular carcinoma and adjacent tissues,and the relationship between Mas receptor and ?-SMA were determined by immunohistochemistry.The hepatic stellate cells with overexpression Mas was constructed by gene transfection,and it was identified by RT PCR.The medium from the hepatic stellate cells with overexpression Mas was added into Huh-7 cells,to detect cell proliferation,cell cycle and apoptosis,using Cell Counting Kit-8 and flow cytometry.Moreover,the ability of migration,invasion and colony-formation viability of Huh-7 cells were detected by Transwell assay and soft-agar colony formation assay.The amounts of TGF ?1 and hydroxyproline in the medium from the hepatic stellate cells with overexpression Mas were measured by ELISA anddigestion method.The effect of tumor formation by the hepatic stellate cells with overexpression Mas was destermined by in vivo tumorigenicity assay.Results Mas and ?-SMA were expressed in human hepatocellular carcinoma and adjacent tissues,and Mas was expressed stronger in adjacent tissues than hepatocellular carcinoma.There was no correlation between the expression of Mas and ?-SMA in hepatocellular carcinoma.The hepatic stellate cells with overexpression Mas was constructed successfully.The hepatic stellate cells could promote the ability of Huh-7 cells proliferation,migration and invasion.While the mediun from Mas overexpression of hepatic stellate cells was added into Huh-7 cells,can weakened apparently the ability of proliferation,migration and invasion of Huh-7,but it was no obvious change detected on cell cycle and apoptosis.Furthermore,there was no obvious change when the amount of TGF?1 and hydroxyproline in the culture medium of the hepatic stellate cells with overexpression Mas.There was also no significance difference in the tumorigenicity assay after co-culture Mas overexpression of hepatic stellate cells and Huh-7 cells.Conclusion Hepatic stellate cells could promote proliferation,migration and invasion of hepatocellular carcinoma cells(Huh-7 cells).Mas overexpression in hepatic stellate cells,could weaken the proliferation,migration,and invasion of hepatoma Huh-7 cells.