【摘要】 目的利用小片段干扰RNA(siRNA)在肝癌细胞内诱导RNA干扰(RNAi),抑制端粒逆转录酶(hTERT)基因表达,在体外进行RNAi对肝癌治疗的试验研究。方法构建针对hTERT基因的siRNA(siRNAhTERT),转导入肝癌细胞7721,在瘤细胞内诱导RNAi,采用流式细胞分析、逆转录聚合酶链反应(RTPCR)法、免疫组化等技术检测siRNA处理前后细胞增殖及hTERT基因表达变化。结果siRNAhTERT对肝癌细胞生长抑制率达37.5%;细胞周期中S期细胞明显减少,G1/G0期细胞显著增加;hTERT基因的mRNA表达由99.4%下调到53.1%,其蛋白表达由86.3%下调到46.6%。结论RNAi在体外明显抑制了肝癌细胞中hTERT基因的表达和瘤细胞增殖。更多还原

【Abstract】 Objective To investigate the hepatocarcinoma therapy by RNA interference, we used siRNA to inhibit hTERT gene. expression in vitro.Methods We constructed a small interference RNA (siRNA) homologous to hTERT gene (siRNA-hTERT), transfected it into hepatocarcinoma cell 7721 to 1induce RNAi, then detected hTERT gene expression in both siRNA-hTERT treatment groups and control groups.Results The expression of hTERT was obviously inhibited by RNAi. The inhibition rate of cell growth was 37.5%. The phase of cell cycle indicated that the S phase decreased, while G1/G0 phase increased. The mRNA level of hTERT was decreased from 99.4% to 53.1%. Its protein expression reduced from 86.3% to 46.6%.Conclusion siRNA-hTERT effectively inhibits hTERT gene expression and cell growth.更多还原