siRNA沉默livin的表达对人结肠癌HT-29细胞增殖、凋亡及侵袭的影响  

Effects of siRNA silencing livin expression on proliferation,apoptosis and invasion of human colon cancer cell line HT-29

作　　者：何文静[1] 黎军和[2] 赵清梅[2] 熊建萍[2] 彭小东[2] 

机构地区：[1]南昌大学第一附属医院输血科,江西南昌330006 [2]南昌大学第一附属医院肿瘤科,江西南昌330006

出　　处：《中国肿瘤生物治疗杂志》2012年第4期392-396,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.30960440)~~

摘　　要：目的:探讨小干扰RNA(small interference RNA,siRNA)沉默人结肠癌HT-29细胞livin表达对HT-29细胞增殖、凋亡和侵袭的影响。方法:合成靶向livin的双链siRNA(livin-siRNA),转染HT-29细胞,RT-PCR及Western blotting检测HT-29细胞中livin mRNA及蛋白的表达,MTT实验检测HT-29细胞的增殖,流式细胞术检测HT-29细胞周期分布及凋亡,细胞侵袭实验检测HT-29细胞侵袭性的变化,caspase-3活性检测试剂盒检测caspase-3活性的变化。结果:Livin-siRNA转染后48 h,与空白组、阴性对照组及脂质体组相比,livin-siRNA转染组HT-29细胞中livin mRNA水平明显下降(0.073±0.007 vs 0.395±0.082、0.423±0.025、0.418±0.032,P<0.05),其蛋白表达也明显下调(0.106±0.003 vs 0.456±0.065、0.473±0.078、0.491±0.045,P<0.05)。转染96 h后,livin-siRNA组HT-29细胞增殖能力明显低于对照组及脂质体组(0.564±0.102 vs0.833±0.127、0.860±0.153,P<0.05),且细胞凋亡率升高[(16.5±2.8)%vs(2.4±0.5)%、(3.7±1.0)%,P<0.05]。侵袭实验显示,livin-siRNA转染后,穿过Matrigel膜的HT-29细胞数量明显少于对照组及脂质体组[(31.3±4.5)vs(101.3±8.6)、(97.4±7.8)个,P<0.05)]。livin-siRNA组HT-29细胞的caspase-3活性低于对照组(0.160±0.023 vs 0.347±0.058,P<0.05)。结论:siRNA沉默livin的表达可抑制HT-29细胞的增殖,诱导细胞凋亡,抑制细胞的侵袭。Objective ： To explore the effects of small interference RNA （siRNA） targeting livin on the proliferation, apoptosis and invasion of human colon cancer cell line HT-29. Methods： Chemically synthetic double-strand siRNA targeting livin （livin-siRNA） was transfected into HT-29 cells, and then RT-PCR and Western blotting were used to detect the expression of livin mRNA and protein in HT-29 cells. MTT assay was performed to analyze the proliferation of HT-29 cells. The cell apoptosis and cell cycle distribution were analyzed by flow cytometry. The invasion assay and caspase-3 detective kit were used to detect the change of invasion and caspase-3 activity in HT-29 cells. Results ： Forty-eight hours after transfection, there was a significant decrease in the expressions of both livin mRNA （ 0. 073 ± 0. 007 vs 0. 395 ± 0.082, 0.423±0.025, 0.418±0.032, P〈0.05） and livin protein （0. 106 ±0.003 vs 0.456 ±0.065, 0.473 ± 0. 078,0. 491 ± 0. 045, P 〈 0.05 ） in the livin-siRNA group, compared with the blank and negative eontrol and liposome groups. Ninety-six hours after transfection, the growth of HT-29 cells in the livin-siRNA group was significantly lower than that in the control and liposome groups （0. 564 ±0. 102 vs 0. 833 ± 0. 127, 0. 860 ± 0. 153, P 〈 0.05 ）, and the rate of apoptosis was obviously increased （ [ 16.5 ± 2.81% vs [ 2.4 ± 0.51%, [ 3.7 ±1.0 ] %, P 〈 0. 05 ）. The invasion assay demonstrated that the number of the migration cells was lower in the livin-siRNA group than in the eontrol and liposome groups （31.3 ± 4.5 vs 101.3 ± 8.6, 97.4 ± 7.8, P 〈 0.05 ）. The aetivity of easpase-3 in the livin-siRNA group was decreased compared with that in the control group （0. 160 ±0. 023 vs 0. 347 ± 0. 058, P 〈0.05）. Conclusion： The siRNA silencing livin expression in HT-29 cells can suppress the proliferation, induce the apoptosis and inhibit the invasion of HT-29 cells.

关 键 词：结肠癌 LIVIN 小干扰RNA 基因治疗 

分 类 号：R735.3[医药卫生—肿瘤] R730.54[医药卫生—临床医学]