联合基因治疗体系pcDNA3.1(-)-shVEGF/yCDglyTK对胃癌细胞增殖的影响  

Combined gene therapy system pcDNA 3.1(-)-shVEGF/yCDglyTK inhibits proliferation of gastric cancer cells

作　　者：陈选民[1] 叶玲[2] 冷爱民[3] 刘霆[3] 

机构地区：[1]南华大学附属第一医院消化科,湖南省衡阳市421001 [2]中南大学湘雅二医院老年病科,湖南省长沙市410008 [3]中南大学湘雅医院消化科,湖南省长沙市410008

出　　处：《世界华人消化杂志》2013年第21期2063-2069,共7页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81272735;教育部博士点新教师基金资助项目;No.200805331090~~

摘　　要：目的:观察由磷酸钙纳米颗粒介导的联合基因治疗体系pcDNA3.1(-)-shVEGF/yCDglyTK对胃癌细胞增殖的影响.方法:以磷酸钙纳米颗粒为载体,介导pcDNA3.1(-)Null,pGenesil-1-hVEGF4-shRNA,pcDNA3.1(-)-CV-yCDglyTK,pcDNA3.1(-)-shVEGF/yCDglyTK转染胃癌细胞SGC7901,RT-PCR检测细胞中yCDglyTK及血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA的表达情况.Western blot分析yCDglyTK及VEGF蛋白的改变.MTT法检测融合基因对转染细胞的杀伤作用.流式细胞术检测转染细胞的凋亡情况.结果:SGC7901细胞转染pcDNA3.1(-)-shVEGF/yCDglyTK质粒后有yCDglyTKmRNA及相关蛋白表达,VEGF表达受抑制达30.3%.相对于转染pcDNA3.1(-)-CV-yCDglyTK质粒组,联合基因体系对胃癌细胞的抑制作用更强(P<0.05).pcDNA3.1(-)-shVEGF/yCDglyTK组细胞凋亡率达到67.9%±4.78%.结论:联合基因治疗体系pcDNA3.1(-)-shVEGF/yCDglyTK较单独的RNA干扰或自杀基因可更有效的杀伤胃癌细胞.AIM： To observe the effect of combined gene ther- apy system pcDNA 3.1（-）-shVEGF/yCDglyTK, which is mediated by calcium phosphate nanoparticles, on the proliferation of human gas- tric cancer cells.METHODS： Expression plasmid vectors pcD-NA3.1（-） NULL, pGenesil-l-hVEGF4-shRNA, pcDNA3.1（-）-CV-yCDglyTK, or pcDNA3.1（-）- shVEGF/yCDglyTK were transfected into human gastric cancer cell line SGC7901. The mRNA and protein expression of yCDglyTK and VEGF was detected by RT-PCR and Western blot, respectively. The proliferation of transfected cells was analyzed by MTT assay. Apoptosis of SGC7901 cells was ana- lyzed by flow cytometry. RESULTS： After transfection with plasmid pcDNA3.1 （-）-shVEGF/yCDglyTK, yCDglyTK mRNA and protein was detectable, and expres- sion of VEGF was decreased by 30.3%. Compared to cells transfected with plasmid pcDNA3.1（-）- CV-yCDglyTK, ceils treated with the combined gene therapy system showed more significant proliferation inhibition （P 〈 0.05） and apoptosis （apoptosis rate： 67.9% ± 4.78%）. CONCLUSION： The combined gene therapy system pcDNA3.1（-）-shVEGF/yCDglyTK has stronger killing effect on gastric cancer cells than RNA interference or suicide gene alone.

关 键 词：联合基因 自杀基因 RNA干扰 血管内皮生长因子 胃癌 

分 类 号：R735.2[医药卫生—肿瘤]