摘要
目的探讨乙型肝炎病毒Ｘ基因对肝细胞恶性变的作用机制。方法将带Ｃ基因、Ｓ基因的载体电转染导入ＨｅＰＧ_２细胞，筛选表达细胞克隆，复苏带Ｘ基因的ＨｅＰＧ_２细胞。ＰＣＲ－ＥＬＩＳＡ检测各株细胞的端粒酶活性。用反义寡核苷酸诱导细胞凋亡，流式细胞仪观测转染了Ｘ基因、Ｃ基因、Ｓ基因细胞的凋亡情况。结果表达细胞克隆经同步化处理，３９．５０％转染Ｘ基因的细胞进入细胞Ｓ周期，其端粒酶活性指数３．９５ ± ０．０７明显高于其它各组细胞。反义寡核苷酸诱导后，转染Ｘ基因细胞凋亡峰明显减小，凋亡率仅１．７５％；其细胞活性与反义寡核苷酸浓度成反比。结论乙型肝炎病毒Ｘ基因上调肝源细胞端粒酶活性，抑制细胞凋亡，这可能是诱导肝细胞恶性变的又一机制。
Objective To observe the effects of hepatitis B virus X gene on hepatic carcinoma. Methods The expression vectors of HBs gene and HBc gene (pCEP4-S and PCEP4-C) were transfected into HepG_2 cells by electroporation technique, and the cells, HepG_2 cells which contained the expression vectors of HBs (pCEP4-X) were recovered and cultured stably. The activity index of telomerase and the apoptosis rates in three cell lines were determined by PCR-ELISA and flow cytometry. Results The 39.50% of cells expressed HBx gene came into S-phase of cell cycle, and the activity index of telomerase was higher in the cell line (3.95 ± 0.07) than in other cell lines and control (F= 20.85, P < 0.01; d' = 2.15, P < 0.05) after synchronized culture. Induced by antisense oligonucleotides against telomerase the apoptosis of cell line expressed HBx gene reduced (apoptosis rate: 1.75%). The activity of cells also reduced with the dose increase of the antisense oligonucleotides and it showed a dose-response relationship. Conclusion The activity of HepG_2 cell can be increased and the cell apoptosis can be reduced by HBx gene. It may be another malignant process of hepatocytes.
出处
《中华肝脏病杂志》
CAS
CSCD
2000年第4期212-214,共3页
Chinese Journal of Hepatology
