Ethanol-induced apoptosis of hepatoma cell line HCC-9204 and its relationship to Bax and Bcl-2 proteins
10.3321/j.issn:1007-8738.2001.04.006
- VernacularTitle:乙醇诱导的肝癌细胞 HCC-9204凋亡及其与 Bax和 Bcl-2蛋白的关系
- Author:
Lianjun YANG
;
Wenliang WANG
;
Xiaohui SI
- Publication Type:Journal Article
- From:
Journal of Cellular and Molecular Immunology
2001;17(4):315-317,358
- CountryChina
- Language:Chinese
-
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.
