摘要
目的探讨microRNA-130b(miR-130b)对胃癌细胞迁移和侵袭能力的影响及作用机制。方法体外培养BGC-823胃癌细胞,通过Lipofectamine2000试剂盒将培养基、miR-130b错义链、miR-130b抑制剂,分别转染进入正常组、对照组、抑制组肿瘤细胞株中。应用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐实验法(MTT)检测转染后胃癌细胞存活率增殖能力变化,划痕实验(wound-healing assay)和侵袭实验(Transwell invasion chamber assay)检测肿瘤细胞迁移、侵袭能力变化,western blot实验检测肿瘤细胞迁移、侵袭相关蛋白分子(Slug、matrix metalloproteinase-9(MMP-9))表达变化情况。结果BGC-823胃癌细胞转染后,与正常组和对照组比较,抑制组细胞增殖能力、迁移和侵袭能力均明显降低(P<0.01)。同时,抑制组细胞迁移、侵袭能力相关蛋白表达水平较正常组和对照组降低(P<0.05)。结论靶向抑制miR-130b可以降低BGC-823胃癌细胞的增值、迁移和侵袭能力。
Objective To investigate the effect of microRNA-130b (miR-130b) on the migration and invasion of gastric cancer ceils and its mechanism.Methods Gastric cancer BGC-823 cells were cultured in vitro and transfected into normal group, control group and inhibition group with medium, miR-130b missense chain and miR-130b inhibitor, respectively using Lipofectamine 2000 Reagent. Gastric cancer cell proliferation ability after transfection was assessed by 3 - ( 4,5 - dimethyl - 2 - thiazolyl ) - 2,5 - diphenyl - 2 - H - tetrazolium bromide (MTT) assay. Cell migration and invasion activity was evaluated by a wound-healing assay and Trans well invasion chamber assay, respectively. Tumor cell migration and invasion activity associated proteins ( slug and matrix metalloproteinase-9 (MMP-9)) expression level were evaluated by western blot assay.Results Gastric cancer BGC-823 cell proliferation ability was significantly reduced compared with normal group and control group (P〈0.01) after transfection. As compared with normal group and control group, migration and invasion activity of inhibition group cell was significantly decreased (P〈 0.01 ). Meanwhile, expression of motility related proteins were significantly decreased compared with normal group and control group (P〈 0.05 ). Conclusions Targeted inhibition of miR-130b can reduce the migration and invasion activity of gastric cancer BGC-823 cells.
作者
陈华
胡海军
单爱军
CHEN Hua(Shenzhen People's Hospital( 2nd Clinical Medical College of Jinan University, Shenzhen, Guangdong, 518020 China)
出处
《齐齐哈尔医学院学报》
2017年第23期2729-2732,共4页
Journal of Qiqihar Medical University
基金
深圳市科技计划项目(JCYJ20170307100359390)
深圳市卫生计生系统科研项目(SZFZ2017073)