RNA干扰沉默血管内皮生长因子对人膀胱癌细胞株T24增殖和凋亡的影响  

Effect of vascular endothelial growth factor knockdown by RNA interference on the growth and apoptosis of bladder carcinoma T24 ceils

作　　者：刘禄成[1] 朱德淳[1] 高瑞娟[2] 郭航[1] 张明[1] 

机构地区：[1]吉林大学第二医院泌尿外科,长春130041 [2]吉林大学基础医学院细胞生物教研室

出　　处：《中华实验外科杂志》2008年第1期74-76,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30571857）

摘　　要：目的观察RNA干扰沉默血管内皮生长因子（VEGF）对膀胱癌细胞株T24增殖和凋亡的影响。方法构建了4个针对VEGF的小干扰RNA（siRNA）表达质粒pGC—siVEGF1-4，脂质体法稳定转染T24细胞。应用逆转录-聚合酶链反应（RT—PCR）和酶联免疫吸附试验（ELISA）检测转染细胞VEGFmRNA和蛋白表达，噻唑蓝（MTT）法测定细胞增殖能力，流式细胞仪检测细胞周期分布和凋亡情况。结果4个VEGF siRNA表达质粒均能明显抑制转染细胞的VEGFmRNA和蛋白表达，使细胞增殖减慢，其中以pGC—siVEGF3最为明显，细胞增殖受抑达38．9％（t=18．49，P〈0．01）。且该组出现了最大程度G0／G1细胞阻滞（59．8±1．4），明显大于空质粒组（48．5±1．5），差异有统计学意义（P〈0．01），该组的细胞凋亡率也最高（17．4％）。结论VEGF siRNA表达质粒可下调T24细胞VEGF表达，抑制膀胱癌细胞生长并促进其凋亡。Objective To observe the effect of vascular endothelial growth factor （VEGF） knockdown by RNA interference on growth and apoptosis of human bladder carcinoma T24 cells. Methods Four recombinant plasmids pGC-siVEGF1-4 targeting VEGF mRNA were constructed, together with a VEGF unrelated control pGC-CONT. The plasmids were transfected into T24 with lipofectin, and positive transfected cell clones were screened with G418. The VEGF mRNA levels were detected by semi-quantitative RT-PCR, and VEGF protein levels in culture media were determined by ELISA. MTT assay was applied to analyze the effect of VEGF siRNA on proliferation of T24 cells. Distributions of cell cycle and cell apoptosis were assessed by flow cytometry. Results RT-PCR and ELISA showed that the levels of VEGF mRNA and protein in the cells of all the four transfected groups were down-regulated compared to the non- transfection cells. The proliferation of all the transfected T24 cells was inhibited, and the effect of pGC- siVEGF3 was the best with the inhibitory rate of 38.9% （ t = 18.49 ,P 〈 0.01 ）. The pGC-siVEGF3 induced a significant G0/Gl arrest （ 59.8 ± 1.4） compared to pGC-CONT control （48.5 ± 1.5, P 〈 0.01 ）, together with a significantly increased apoptosis rate （ 17.4% ）. Conclusion The recombinant siRNA expression plasmid targeting VEGF can inhibit cell proliferation and induce cell apoptosis in T24 cells through significantly interfering VEGF mRNA and protein expression.

关 键 词：膀胱癌 RNA干扰 血管内皮生长因子 增殖 脱噬作用 

分 类 号：R737.14[医药卫生—肿瘤] R735.2[医药卫生—临床医学]