DNA载体途径的RNA干扰技术对前列腺癌细胞系ALVA-41端粒酶活性的抑制作用

RNA interference directed by small interference RNA of hTERT expressed in prostate carcinoma cell ALVA-41

目的:探讨DNA载体途径的RNA干扰技术抑制肿瘤细胞的作用和机制。方法:设计和构建由U6启动子驱动产生hTERT的siRNA表达质粒,转染至ALVA-41前列腺肿瘤细胞,以TRAP-ELISA方法观察细胞端粒酶活性,W estern-b lot检测端粒酶蛋白的表达,细胞记数法检测对细胞生长的影响。结果:构建的针对hTERT基因的U6表达质粒具有明显的干扰效果,受到特异hTERT-siRNA干扰的ALVA-41肿瘤细胞端粒酶表达下调,细胞生长受抑。结论:应用RNA干扰技术可阻止hTERT基因的表达,抑制肿瘤细胞的增殖。

Objective： To develop a DNA vector-based RNA interference technology that inhibite the activity of the telomerase of the carcinoma cell in order to suppress the preliforation of the cells. Methods： A plasmid including U6 promoter and siRNA of hTERT was designed and constructed. The plasmid was transfected into ALVA-41 cell line. The telomerase activity was tested by telomerase repeat amplification protocol ELISA （TRAP-ELISA）. Cell calculating assay was used to assay the cell proliferation activity, and hTERT expression was assessed by Western blot. Results：The U6 expression plasmid that was constructed for hTERT showed obvious interfering effect, hTERT-siRNA could down regulate the expression of hTERT protein, inhibit telomerase activity and proliferation of ALVA-41 cells. Conclusion： siRNA of hTERT can inhibit the expression of human telomerase and proliferation of ALVA-41 cells.