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作 者:曾春林[1] 赵亚力[1] 高江平[2] 韩为东[1] 李琦[1]
机构地区:[1]解放军总医院基础所分子生物学研究室,北京100853 [2]解放军总医院泌尿外科,北京100853
出 处:《军医进修学院学报》2006年第1期37-39,共3页Academic Journal of Pla Postgraduate Medical School
基 金:北京自然科学基金(7042059)
摘 要: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.
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