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

发布时间：2018-03-14 06:04

本文选题：胃肿瘤　切入点：miR-　出处：《实用医学杂志》2017年05期 　论文类型：期刊论文

【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(P0.05)。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(P0.05)。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。
[Abstract]:Objective: to construct a recombinant AGS cell line with stable and high expression of miR-126, and to study the effect of miR-126 on the proliferation and metastasis of gastric cancer cell line in vitro. Methods: AGS cell line infected with lentivirus Lv-has-miR-126 was constructed and the difference of miR-126 expression was detected by qRT-PCR. After the successful construction of the recombinant cell line with stable and high expression of miR-126, the changes of cell proliferation ability in vitro were detected by CCK-8 and plate clone formation assay and the migration in vitro was detected by transwell migration assay and invasion assay, respectively. Invasiveness. Results: recombinant cell lines were successfully constructed, The expression level of miR-126 in recombinant cells was significantly higher than that in control cells by green fluorescence RT-PCR. The results showed that there was no significant difference in the growth and clone formation of gastric cancer cells after up-regulation of miR-126. The migration and invasion ability of the cells in the high expression group of miR-126 was significantly higher than that in the control group (P 0.05). Conclusion: the recombinant AGS cell line with stable and high expression of miR-126 was successfully constructed. It is preliminarily confirmed that miR-126 can promote the migration and invasion of gastric cancer AGS cells, suggesting that miR-126 may be closely related to gastric cancer metastasis.
【作者单位】：南方医科大学南方医院检验科;
【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)
【分类号】：R735.2

【相似文献】