论文部分内容阅读
目的构建针对端粒酶hTERT的双H1启动子SECs,并探讨其转录生成的siRNA对HepG2细胞端粒酶活性的抑制作用。方法利用融合PCR技术,构建2个针对人端粒酶hTERT基因外显子不同片断的双H1启动子SECs,分别转染人肝癌细胞HepG2,转录生成siRNAs。用TRAP法和PCR-EIA法检测端粒酶活性,分析双H1启动子SECs对HepG2细胞端粒酶表达的干扰作用。结果成功构建针对hTERT的双H1启动子SECs,其转录产物siRNA可明显抑制HepG2细胞端粒酶的活性。结论siRNA SECs对HepG2细胞端粒酶hTERT基因表达有明显的干扰作用,为临床开展对肿瘤端粒酶基因干扰抑制的实验研究奠定了基础。
OBJECTIVE: To construct double H1 promoter SECs targeting telomerase hTERT and investigate the inhibitory effect of its transcribed siRNA on telomerase activity in HepG2 cells. Methods Two double-promoter H1s targeting different segments of human telomerase hTERT gene were constructed by fusion PCR and transfected into HepG2 cells to generate siRNAs. The telomerase activity was detected by TRAP and PCR-EIA, and the interference of double H1 promoter SECs on telomerase expression in HepG2 cells was analyzed. Results Double-H1 promoter SECs targeting hTERT were successfully constructed and the transcripts of siRNAs could inhibit the activity of telomerase in HepG2 cells. Conclusion siRNA SECs can significantly inhibit the expression of telomerase hTERT gene in HepG2 cells, which lays the foundation for the clinical research on the inhibition of telomerase gene interference.