Ethanol-induced apoptosis of hepatoma cell line HCC-9204 and its relationship to Bax and Bcl-2 proteins / 细胞与分子免疫学杂志

ABSTRACT

Aim To explore the ethanol-induced apoptosis effect on hepatocellular carcinoma(HCC) cells and its relationship to the expression of apoptosis associated genes, bax and bcl-2. Methods The cytotoxic effect of 20～ 100 mL/ L ethanol on HCC cell line HCC-9204 was tested by thiazolyl-blue (MTT) assay. Then apoptosis of HCC-9204 cells was induced with 60 mL/ L of ethanol for 6 h. The morphological change, DNA breakage and the change of DNA content of different cell cycles of the apoptotic cells were detected by May-Grunwald Giemsa(MGG) staining, and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay and flow cytometer respectively. The changes of expression level of Bax and Bcl-2 proteins were detected by immunocytochemical staining and image analysis. Results The higher the concentration of ethanol was, the stronger the cytotoxic effect on HCC-9204 cells was. 60 mL/ L of ethanol could lead to obviously morphological apoptotic changes of HCC-9204 cells, and majority of the cells were TUNEL positive by TUNEL labeling assay. Typical apoptotic sub G1 peak was observed by flow cytometer. The level of Bax protein expression increased significantly after induced with 60 mL/ L of ethanol for 6 h, no expression of Bcl 2 were found before and after induced with ethanol. Conclusion Low dose of ethanol can induce apoptosis of HCC-9204 cells obviously, and occurance of the apoptosis is related to the increase of the level of Bax protein expression.