RNAi抑制hTERT基因治疗乳腺癌的实验研究

Experimental research of targeting hTERT gene inhibited in breast cancer therapy by RNA interference

目的利用siRNA在乳腺癌细胞内诱导RN Ai,抑制hTERT基因表达,在体外细胞水平探讨RNAi对乳腺癌治疗的可行性。方法构建靶向hTERT基因的siRNA(hTERT-siRNA),转导入乳腺癌MCF-7细胞,在瘤细胞内诱导RNAi,采用细胞增殖抑制实验、RT-PCR法、Western blot等技术检测siRNA处理前后瘤细胞增殖及hTERT基因表达变化。结果 hTERT-siRNA对乳腺癌细胞生长抑制率达43%;hTERT基因的mRNA表达明显降低,蛋白表达平均下调了39.8%。结论 siRNA在体外明显抑制了乳腺癌细胞中hTERT基因的表达和瘤细胞增殖。

Objective To investigate the feasibility and specificity of breast cancer gene therapy, we utilized RNAi to inhibit hTERT gene expression in MCF-7 cells of breast cancer. Methods Small intererence RNA （siRNA） homologous to hTERT gene （hTERT-siRNA） was recombined with pGenesil-1 vector, then transfected it into MCF-7 cells of breast cancer to induce RNAi, and then detected hTERT gene ex- pression alterations and tumor cell proliferation in both hTERT-siRNA treatment groups and control groups by MTT assay, RT-PCR and Western blot. Results The expression of hTERT had been obviously inhibited by siRNA. The inhibition rate of cell proliferation was 43%. The mRNA level of hTERT was obviously de- creased, and its protein expression reduced by 39.8%. Conclusions siRNA-hTERT effectively inhibited hTERT gene expression and breast cancer cell proliferation in vitro.