Transfection of apoptosis related gene Fas ligand in human hepatocellular carcinoma cells and its significance in apoptosis.
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
Levels of soluble Fas ligand (sFasL) in a group of patients with hepatitis B virus (HBV)-induced chronic hepatitis, HBV-positive liver cirrhosis and HCC were evaluated. In a further study, 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 was detected by immunohistochemistry. After being stained by annexin V and propidium iodine, cells were passed through a flow cytometer and examined by a fluorescence microscope and a laser scanning microscope.


RESULTS
The sFasL levels were significantly lower in patients with HCC when compared to the patients with hepatitis or liver cirrhosis. In comparison with untransfected cells, the soluble FasL could be detected in the supernatant of transfected cells. FasL was expressed on the membranes and cytoplasm of transfected cells. The apoptotic cell rate was 36.30% in transfected cells, and was 11.53% in untransfected cells. Moreover, the different stage of apoptotic cells could be distinguished by annexin V and propidium iodine staining.


CONCLUSIONS
Fas ligand is an apoptotic pathway of HCC cells.
Chen
Jun
J
Department of Infectious Diseases, Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China. drchenjun@hotmail.com
Su
Xian-Shi
XS
Jiang
Yong-Fang
YF
Gong
Guo-Zhong
GZ
Zheng
Yu-Huang
YH
Li
Gui-Yuan
GY
D017209:Apoptosis / Q000502:physiology*; D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000503:physiopathology*; D045744:Cell Line, Tumor; D053222:Fas Ligand Protein; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology / Q000503:physiopathology*; D008562:Membrane Glycoproteins / Q000235:genetics*; D014162:Transfection
C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins