三氧化二砷对肝癌细胞survivin基因和端粒酶活性的影响

Influence of arsenic trioxide onsurvivingene expression and telomerase activity of hepatoma cell lines

目的：探讨三氧化二砷（As2O3）诱导肝癌细胞凋亡时survivin基因表达和端粒酶活性的变化。方法：采用倒置相差显微镜观察细胞形态，流式细胞仪分析细胞亚二倍体百分率，逆转录-多聚酶链式反应检测细胞survivin mRNA表达，免疫组织化学染色检测细胞survivin蛋白的表达，端粒酶检测试剂盒检测端粒酶活性。结果：0．25～2．00μmol／L As2O3明显抑制肝癌Bel-7402细胞生长，诱导细胞凋亡，survivin mRNA和蛋白表达上调，端粒酶活性下降，呈时间、剂量依赖关系。结论：As2O3抑制肝癌Bel-7402细胞增殖，诱导细胞凋亡，可能与其降低端粒酶活性有关；而survivin基因表达上调，可能是抵抗As2O3诱导凋亡的途径之一。

Objective：To explore the changes of survivin gene expression and telomerase activity during apoptosis of hepatoma cell lines induced by arsenic trioxide. Methods：Cell morphological changes were observed under phase contrast microscope. Percentage of diploid cells was analyzed by flow cytometry. The expression level of survivin mRNA was measured by reverse transcription polymerase chain reaction （RT-PCR） and survivin protein level was determined by immunohistochemistry. The telomerase activities were detected using the quantitative telomerase PCR ELISAPLUS K kit. Results.. Arsenic trioxide 0. 25-2.00 μmol/L markedly inhibited growth of hepatoma Bel-7402 cells, induced apoptosis, up-regulated expression of survivin mRNA and protein, reduced telomerase activity in a time- and dose-dependent manner. Conclusion：Arsenic trioxide significantly inhibited the growth of Bel-7402 cells and induced apoptosis, which may be correlated with decrease in telomerase activity. Up-regulation of survivin gene expression was one of the factors that resist arsenic trioxide-induced apoptosis.