hTERT siRNA表达载体的构建及对转染的HeLa细胞生长抑制作用

Construction of siRNA expression vectors for human telomerase reverse transcriptase and their inhibitory effect on transfected HeLa cells in vitro

目的:通过RNA干涉技术抑制肿瘤细胞端粒酶表达,探讨干涉后对肿瘤细胞生长的抑制作用.方法:根据人端粒酶逆转录酶(hTERT)mRNA编码序列,设计RNA干涉靶点,构建siRNA(smallinterferencingRNA)表达载体,并转染HeLa细胞,通过RT PCR法观察重组质粒转染肿瘤细胞后端粒酶mRNA、蛋白含量及细胞生长情况.结果:构建的siRNA表达载体可以使HeLa细胞端粒酶逆转录酶mRNA及其蛋白含量降低,转染质粒的细胞生长增殖速度减慢.结论:成功构建了针对人端粒酶逆转录酶的siRNA表达载体,通过转染HeLa细胞,可有效抑制细胞中端粒酶的表达,并引起细胞生长抑制.

AIM: To study the growth inhibition of tumor cells by inhibiting telomerase expression of tumor cells with RNA interference (RNAi) technique. METHODS: According to the encoding sequence of mRNA of hTERT (human telomerase reverse transciptase), the target site for RNAi technique was designed and the vector for siRNA (small interfering RNA) expression in tumor cells was constructed. The HeLa cell line was transfected and the inhibition of the expression of telomerase was further observed. RESULTS: The constructed vector significantly inhibited the expression of mRNA and protein of telomerase in HeLa cells. The growth of the transfected cells was inhibited. CONCLUSION: siRNA expression vectors targeting hTERT are successfully constructed. After transfection of the constructed vectors into HeLa cells, the expression of telomerase in HeLa cells is effectively inhibited and thus slows down the cell growth of the HeLa cells.