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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 investigate the mechanism of hepatitis B virus X gene on hepatocellular malignant transformation. Methods HepG2 cells were transfected with the vector carrying C gene and S gene. The cloned cells were recovered and the HepG2 cells with X gene were recovered. The telomerase activity of each cell line was detected by PCR-ELISA. Antisense oligodeoxynucleotides were used to induce apoptosis. Flow cytometry was used to observe the apoptosis of X gene, C gene and S gene. Results The cell clones were synchronized and the percentage of telomerase activity index was 3.95 ± 0.07 in 39.50% of cells transfected with X gene. The telomerase activity index was significantly higher than that in other groups. After induction with antisense oligonucleotide, the apoptotic peak of the transfected X gene was significantly reduced, with a apoptotic rate of only 1.75%. The cell activity was inversely proportional to the concentration of antisense oligonucleotide. Conclusion Hepatitis B virus X gene upregulates telomerase activity and inhibits apoptosis in hepatocytes, which may be another mechanism of hepatocellular malignant transformation.