目的

利用siRNA在乳腺癌细胞内诱导RNAi，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 expression 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 decreased， and its protein expression reduced by 39.8%.

Conclusions

siRNA-hTERT effectively inhibited hTERT gene expression and breast cancer cell proliferation in vitro.