siRNA沉默Livin基因对胃癌细胞生长、凋亡的影响  

Effects of Livin gene silencing by siRNA on apoptosis and proliferation of gastric cancer cells

作　　者：陈鹏[1] 孔令斌[2] 王春松[1] 杨景玉[3] 

机构地区：[1]山东大学医学院,山东省济南市275012 [2]济宁医学院,山东省济宁市272013 [3]山东省寄生虫病防治所,山东省济宁市272033

出　　处：《世界华人消化杂志》2009年第13期1346-1349,共4页World Chinese Journal of Digestology

基　　金：山东省教育厅基金资助项目;No.J08LH03~~

摘　　要：目的:观察小分子干扰RNA(siRNA)沉默Livin基因在胃癌BGC-823细胞中的表达,并探讨Livin基因对胃癌细胞生长、凋亡的影响.方法:自行设计两条针对Livin基因的siRNA:Livin-sh-1和Livin-sh-2,以此构建相应的表达载体并分别转染至对数生长期胃癌BGC-823细胞,经G418筛选后分别采用半定量RT-PCR检测不同siRNA实验分组细胞BGC-823mRNA水平变化,四氮唑盐比色法(MTT)检测细胞增殖、流式细胞仪检测胃癌细胞的凋亡.结果:siRNA对照组与空siRNA载体组Livinα/βmRNA表达差别无显著性;但转染siRNA组Livinα/βmRNA表达显著低于空白对照组和空siRNA载体组(Livinα:0.11±0.07vs0.37±0.10,0.34±0.08;Livinβ:0.13±0.04vs0.43±0.09,0.45±0.11,均P<0.05).空白对照组与空siRNA载体组相比,24、48、96h和1wk时细胞生长未受影响;而siRNA组在转染后24h和48h细胞生长未受影响,但在96h和1wk时则被明显抑制(P<0.01).转染siRNA组的细胞的凋亡率与空白对照组和转染空siRNA载体组相比显著增加(14.85%±1.35%vs4.51%±0.36%,6.13%±0.71%,均P<0.05).结论:siRNA沉默Livin基因能抑制胃癌细胞的生长,促进胃癌细胞的凋亡,Livin基因有可能成为胃癌治疗的新靶点.AIM： To investigate the expression and effect of Livin gene silencing by siRNA on growth and apoptosis in gastric cancer BGC-823 cells. METHODS： Two siRNAs （Livin-sh-1 and Livinsh-2） were self-designed as expression vector and transfected into gastric cancer BGC-823 cells. After G418 positive clone selection, BGC-823 mRNA was measured by semi-quantitative RTPCR, cell proliferation detected by MTT, and gastric cell apoptosis by flow cytometry. RESULTS： Compared with cells without siRNA transfection, expression of Livin α/β mRNA was decreased significantly in siRNA control group （Livin α： 0.11 ± 0.07 vs 0.37 ± 0.10, 0.34 ±0.08; Livin β： 0.13 ± 0.04 vs 0.43 ± 0.09, 0.45 ± 0.11, all P 〈 0.05）. Compared with empty siRNA vector group, cell growth at 24 h, 48 h, 96 h, and 1 wk was not affected in blank control group. Cell growth in siRNA group was not significantly affected at 24 h and 48 h, but was significantly inhibited at 96 h and 1wk （P 〈 0.01）. Cell apoptosis was significantly higher in siRNA in transfection group than in non-transfected group or than in empty siKNA vector transfection group （14.85% ± 1.35% vs 4.51% ± 0.36%, 6.13% ± 0.71%, both P 〈 0.05）. CONCLUSION： Livin gene silenced by siRNA induces growth suppression and apoptosis of gastric cancer BGC-823 ceils. Livin gene stands a chance as a new target for gastric cancer treatment.

关 键 词：小分子干扰RNA LIVIN基因 胃癌 

分 类 号：R735.2[医药卫生—肿瘤]