慢病毒介导稳定过表达ADP核糖化因子鸟苷酸激酶1可促进胃癌细胞株SGC7901的增殖和迁移  

Lentivirus-mediated over-expression of ADP ribosylation factor guanylate kinase 1 accelerates proliferation and migration of gastric carcinoma SGC7901 cells

作　　者：范永刚[1] 王伟[1] 姚国良[1] 封冰[1] 张贝克[1] 解刚强[1] 

机构地区：[1]河南科技大学第一附属医院新区医院普外科,洛阳471003

出　　处：《中华实验外科杂志》2015年第5期1045-1047,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察ADP核糖化因子鸟苷酸激酶1（ASAP1）对胃癌细胞株SGC7901增殖、迁移能力的影响.方法 构建靶向过表达ASAP1基因的慢病毒载体LV-ASAP1-GFP,感染胃癌SGC7901细胞株,经过筛选,建立稳定过表达ASAP1基因的胃癌SGC7901细胞株.通过实时定量聚合酶链反应（Real-time PCR）、Western blot分别检测ASAP1基因mRNA与蛋白表达量,噻唑蓝（MTT）比色法、Transwell体外细胞迁移实验分别研究稳定过表达ASAP1基因对细胞增殖和迁移的影响.结果 成功包装靶向过表达ASAP1基因的慢病毒载体LV-ASAP1-GFP稳定过表达ASAP1基因的人胃癌SGC7901细胞.与空载对照LV-GFP组和阴性SGC7901细胞组比较,Real-time PCR与Western blot证实,慢病毒LV-ASAP1-GFP组mRNA相对表达量为对照组的255.4％,蛋白相对表达量为对照组的290.2％;MTT比色法显示LV-ASAP1-GFP组细胞增殖能力增强,在1、2、3d后重复检测,分别达到了同时间空白对照组的130.1％、136.0％和149.1％ （P ＜0.05）;细胞迁移实验显示LV-ASAP1-GFP组细胞迁移能力显著增强,穿膜细胞数为对照组的230.3％ （P＜0.01）.结论 慢病毒介导稳定过表达ASAP1基因能使胃癌SGC7901细胞株的增殖能力及迁移能力增强.Objective To construct a stable over-expression lentivirus-mediated vector and transfect it into human gastric cancer cell line SGC7901 for investigating its effects on proliferation and migration of GC7901 cells.Methods ADP ribosylation factor guanylate kinase 1 （ASAP1） gene coding region was cloned into lentivirus vector.Lentivirus particles were infected into the human gastric carcinoma cell line SGC7901 to upregulate the expression of ASAP1 gene.The up-regulated efficiency of targeting ASAP1 gene at mRNA level was detected by real-time quantitative polymerase chain reaction （Real-time PCR）,the effect on proliferation of mesenchmal stem cells was assayed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay,and the migration ability was detected by Transwell motility assay.Results The lentiviral vector targeting ASAP1 gene was constructed successfully,and a stable human gastric cancer cell line SGC7901 line that up-regulated ASAP1 was established.Real-time quantitative PCR and Western blotting results showed that the expression of ASAP1 gene was efficiently up-regulated by infecting LV-ASAP1-GFP （P 〈 0.01）.255.4% of control group on mRNA expression and 290.2% of control group on protein expression.The 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay and growth curve showed that the over-expression of the ASAP1 gene successfully increased the proliferative capability of SGC7901 cells,with optical density values reaching 130.1%,136.0% and 149.1% of control group at 1,2,3 d respectively.The Transwell assay also showed similar increasing results on the migration ability （P 〈 0.01）.Conclusion The lenvivirus-mediated over-expression of the ASAP1 gene increases the proliferation and migration abilities of human gastric cancer cell line GC7901.

关 键 词：胃癌 ADP核糖化因子鸟苷酸激酶1 

分 类 号：R735.2[医药卫生—肿瘤]