南方医科大学学报2009，Vol.29Issue(11)：2187-2190,4.
靶向hTERT基因表达载体的构建和其对人乳腺癌细胞端粒酶活性及细胞增殖的影响
Construction of an expression vector carrying short hairpin RNA targeting hTERT gene and its effects on breast cancer cell telomerase activity and proliferation in vivo
摘要
Abstract
Objective To construct a RNA interference expression vector targeting human telomerase reverse transcriptase gene (hTERT) gene and investigate its effects on telomerase activity and proliferation in breast cancer cells in vitro. Methods The shRNA sequences targeting hTERT gene were designed and recombined into pSuper-retro-puro vector. The breast cancer cell lines MCF-7 and MDA-MB231 were transfected with the recombined vector, and the telomerase activity of the cells was tested by telomerase repeat sequence amplification-enzyme linked immunosorbent assay (TRAP-ELISA). The proliferation of the transfected cells was assessed using MTT and soft-agar clone formation assays. Results The recombinant plasmids pSuper-retro-puro-TERT RNAi#1 and #2 were successfully constructed as confirmed by enzymatic digestion and DNA sequencing. The telomerase activity in the transfected breast cancer cells were down-regulated significantly as compared with that in negative control cells (P<0.005). The transfection resulted in significant inhibition of the proliferation of both MCF-7 and MDA-MB231 cells as detected by MTT assay (P<0.05) and soft agar clone formation assay (P<0.001). Conclusion Transfection with the recombinant plasmid containing the shRNA targeting hTERT gene can down-regulate telomerase activity and inhibit proliferation of breast cancer cells in vitro, suggesting the potential of gene therapy targeting telomerase in the treatment of breast cancer.关键词
乳腺癌/端粒酶/hTERT/RNA干扰/基因治疗Key words
breast cancer/ telomerase/ human telomerase reverse transcriptase gene/ RNA interference/ gene therapy分类
医药卫生引用本文复制引用
刘祥厦,姚陈,张辉,王三明,王深明..靶向hTERT基因表达载体的构建和其对人乳腺癌细胞端粒酶活性及细胞增殖的影响[J].南方医科大学学报,2009,29(11):2187-2190,4.基金项目
广东省自然科学基金(021907) （021907）
