慢病毒载体介导的miR-126稳定高表达促进胃癌AGS细胞的迁移和侵袭  

Lentiviral vector mediated up-regulation of miR-126 promotes migration and invasion on AGS cells

作　　者：殷小凤 李月[1] 李欣[1] 李海侠[1] 裘宇容[1] 

机构地区：[1]南方医科大学南方医院检验科,广州市510515

出　　处：《实用医学杂志》2017年第5期696-700,共5页The Journal of Practical Medicine

基　　金：广东省自然基金项目(编号:2016A030313525);广州市科技计划项目(编号:201607010015)

摘　　要：目的:构建稳定高表达miR-126的胃癌AGS重组细胞系,并研究miR-126在体外对该胃癌细胞系增殖、转移能力的影响。方法:构建慢病毒Lv-has-miR-126感染AGS细胞系,经qRT-PCR检测miR-126表达差异,确认miR-126稳定高表达重组细胞系构建成功后,应用CCK-8和平板克隆形成实验检测细胞体外增殖能力变化;Transwell迁移及侵袭实验检测细胞在体外的迁移、侵袭能力。结果:重组细胞系构建成功,绿色荧光表达良好,qRT-PCR证实重组细胞miR-126的表达水平较对照细胞显著增高(P<0.05)。CCK-8增殖及克隆形成实验显示上调miR-126后胃癌细胞生长及克隆形成数无明显差异;而在迁移及侵袭实验中miR-126高表达组细胞迁移及侵袭能力显著高于对照组(P<0.05)。结论:成功构建了稳定高表达miR-126的AGS重组细胞系,初步证实了miR-126具有促进胃癌AGS细胞迁移与侵袭的作用,提示miR-126可能与胃癌的转移密切相关。Objective To establish an AGS cell line that stably expressing miR-126 and to study the effect of miR-126 on the proliferation and metastatic abilities of the AGS cell line in vitro. Methods AGS cells were infected by lentivirus with Lv-has-mir-126. After confirmation by RT-PCR, CCK-8 and clone formation assays were used to evaluate the effect of miR- 126 on AGS cell growth. Transwell migration and invasion assays were used to evaluate the effect of miR- 126 on metastasis of AGS cells. Results We verified correct construction of recombinant AGS cells. RT-PCR confirmed mRNA levels of miR- 126 existed significantly differences among the recombinant cell lines （P 〈 0.05）. Proliferation assays and clone formation assays did not show a remarkable growth suppression in AGS-mir-126 cell line. However, transwell assay showed a notable acceleration in AGS-mir- 126 （P 〈 0.05）. Conclusions We successfully constructed recombinant AGS cell line with stably high miR-126 expression level. MiR-126 could facilitate the metastasis of AGS cell in vitro.

关 键 词：胃肿瘤 MIR-126 转移 侵袭 慢病毒 稳定表达 

分 类 号：R735.2[医药卫生—肿瘤]