The biological effect of Fas Ligand transfected HepG2 cell / 中国免疫学杂志

ABSTRACT

Objective:To evaluate the expression of apoptosis related gene Fas Ligand(FasL) in human hepatocellular carcinoma(HCC) cells HepG2 and its significance in apoptosis.Methods:The recombinant eukaryotic expression plasmid pcDNA3.1hisB- FasL was transfected into HCC cells HepG2 by lipofection, and then soluble FasL was examined in the supernatant of culture cells by EIA, FasL expression in HepG2 cells were detected by immune histochemistry. After stained by annexin V and propidium iodine, cells were passed throw flow cytometer and examined by fluorescence microscope and sym-focus laser scaning microscope.Results:In comparison with untransfected cells,the soluble FasL could be detected in the supernatant of transfected cells, Fas L can be expressed in the membrane and cytoplasm of transfected cells. The apoptotic cell rate in transfected cells was 36.30%, as the control, untransfected cells was 11.53%.Moreover, the different stage of apoptotic cells could be distinguished by annexin V and propidium iodine stain.Conclusion:This supports a novel pathway of HCC cells were apoptotic itself via the CD95-CD95 ligand system without involvement of immune cells.