VEGF基因表达抑制对胃腺癌细胞SGC-7901增殖的影响  

Effect of down-regulating vascular endothelial growth factor gene on proliferation of gastric cancer cell line SGC-7901

作　　者：徐文华[1] 葛银林[1] 徐宏伟[1] 王秀丽[1] 耿芳宋[1] 

机构地区：[1]青岛大学医学院生物化学与分子生物学教研室,山东省青岛市266021

出　　处：《世界华人消化杂志》2006年第7期655-659,共5页World Chinese Journal of Digestology

基　　金：青岛市科技局科技计划项目;03-1-NY-14

摘　　要：目的:研究siRNA(small interfering RNA)对人胃腺癌细胞SGC-7901的VEGF基因表达的影响．方法:选择血管内皮生长因子(VEGF)基因为靶基因,设计两组针对VEGF mRNA的小干扰RNA,合成DNA寡核苷酸链,体外转录合成siRNA．以人胃腺癌细胞系SGC-7901为靶细胞,应用脂质体转染的方法,将siRNA导入细胞．采用Hoechst33258染色观察siRNA作用于SGC-7901细胞中出现凋亡小体的情况．流式细胞仪检测细胞周期的改变,RT-PCR法比较转染前后VEGF mRNA表达水平的变化, ELISA法检测细胞培养液中VEGF蛋白分泌量的变化．结果:两组siRNA转染后均能有效地抑制 SGC-7901细胞的生长,诱导细胞凋亡产生凋亡小体．siRNA作用于SGC-7901细胞．其细胞周期均发生了明显的变化,主要表现为G0／G1 期细胞增多,S期细胞减少,并使细胞周期阻滞于G0／G1期(siRNA1组,siRNA2组G0／G1期VS 对照组G0／G1期:75．04％,76．52％ vs 58．37％, P<0．01;siRNA1组,siRNA2组S期vs对照组S 期:17．82％,16．73％ vs 39．52％,P<0．01),而其他组则无明显变化．VEGF mRNA的表达量大幅度减少(siRNA1组,siRNA2组vs对照组: 0．638±0．078,0．656±0．085 vs 0．941±0．046, P<0．01),相对应的VEGF蛋白水平也显著降低(164．7±22．7,166．3±26．6 vs 414．0±61．5, P<0．01),而其他组siRNA转染后则无上述作用．结论:应用靶向VEGF基因RNA干扰技术可以有效抑制胃癌细胞SGC-7901的增殖．AIM： To study the effect of small interfering RNA （siRNA） on the gene expression of vascular endothelial growth factor （VEGF） as well as the proliferation of gastric cancer cell line SGC-7901. METHODS： Two groups of siRNA were designed targeting VEGF mRNA and finally obtained by in vitro transcription. Then the obtained siRNA was transfected into cultured human gastric cancer cell line SGC-7901 by Lipofectamine. Hoechst33258 staining was used to observe the apoptotic bodies in the cells, and flow cytometry was used to detect the change of cell cycles. The level of VEGF mRNA expression was analyzed by reverse transcription polymerase chain reaction （RT-PCR） and the secretion of VEGF protein in the supematant was examined by enzyme-linked immunosorbent assay （ELISA）. RESULTS： Both groups of VEGF-targeted siRNA inhibited the proliferation of SGC-7901 cells and induced cell apoptosis effectively and after transfection. The percentage of G0/G1 phase cells was significantly increased in siRNA1- and siRNA2-transfected cells in comparison with that in the control cells （75.04%, 76.52% vs 58.37%, P 〈 0.01）, but the percentage of S phase cells was significantly decreased （17.82%, 16.73% vs 39.52%, P 〈 0.01）. The level of VEGF mRNA expression was significantly inhibited in siRNA1- and siRNA2-transfected cells as compared with that in the controls （0.638 ± 0.078, 0.656 ± 0.085 vs 0.941 ± 0.046, P 〈 0.01）, and the secretion of VEGF protein was also notably decreased in both siRNA-transfected cells （164.7 ± 22.7, 166.3 ± 26.6 vs 414.0 ± 61.5, P 〈 0.01）. CONCLUSION： VEGF-targeted siRNA can effectively inhibit the proliferation of human gastric cancer cell line SGC-7901.

关 键 词：胃癌 SGC-7901 RNA干扰 血管内皮细胞 生长因子 VEGF基因表达抑制 血管内皮生长因子 

分 类 号：R735.2[医药卫生—肿瘤]