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[微小RNA-191-5p通过靶向细胞周期蛋白依赖性激酶6抑制胃癌细胞生长]

[miRNA-191-5p represses cell growth by targeting CDK6 in gastric cancer].

作者信息

Cao N J, Hou H H, Li F, Guo S T, Wang Y

机构信息

Department of Biochemistry and Molecular Biology, Basic Medical College, Shanxi Medical University, Taiyuan 030001, China.

Shanxi Cancer Hospital, Taiyuan 030013, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2020 Dec 15;100(46):3689-3693. doi: 10.3760/cma.j.cn112137-20200407-01111.

Abstract

To investigate the effects of miR-191-5p on cell migration, clone formation and proliferation of gastric cancer (GC) cells. The level of miR-191-5p expression was detected by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in 60 paired GC tissues and their adjacent normal tissues. miR-191-5p overexpression was achieved by transfection of construct pcDNA-miR-191-5p into GC cells. The migration, clone formation and proliferation of GC cells were detected by the scratch wound assay, clone formation assay and cell counting kit-8 (CCK-8), respectively. Low expression of miR-191-5p was achieved with miRNA-191-5p inhibitor. The binding sites of cyclin-dependent kinase 6 (CDK6) and miR-191-5p were analyzed using TargetScan software, and the interaction of CDK6 and miR-191-5p was verified using dual-fluorescence reporter gene expression. Western blot (WB) was used to detect the effect of miR-191-5p on the expression of p21 and CDK6 proteins. miR-191-5p decreased in 53 cases (88%) of GC tissues compared to their controls. Furthermore, overexpression of miR-191-5p effectively inhibited the migration, clone formation and proliferation of GC cells (<0.05). Dual-fluorescence reporter confirmed that miR-191-5p bound to 3'UTR of CDK6. WB showed that pcDNA-miR-191-5p inhibited the CDK6 expression but promoted the p21. Down-regulation of miR-191-5p has a correlation with the progression of GC. Overexpression of miR-191-5p can decrease the expression of CDK6 and inhibit the growth of GC cells.

摘要

探讨miR-191-5p对胃癌(GC)细胞迁移、克隆形成及增殖的影响。采用实时逆转录聚合酶链反应(RT-PCR)检测60对GC组织及其癌旁正常组织中miR-191-5p的表达水平。通过将构建体pcDNA-miR-191-5p转染至GC细胞实现miR-191-5p过表达。分别采用划痕试验、克隆形成试验和细胞计数试剂盒-8(CCK-8)检测GC细胞的迁移、克隆形成及增殖情况。使用miRNA-191-5p抑制剂实现miR-191-5p的低表达。利用TargetScan软件分析细胞周期蛋白依赖性激酶6(CDK6)与miR-191-5p的结合位点,并通过双荧光报告基因表达验证CDK6与miR-191-5p的相互作用。采用蛋白质免疫印迹法(WB)检测miR-191-5p对p21和CDK6蛋白表达的影响。与对照相比,53例(88%)GC组织中miR-191-5p表达降低。此外,miR-191-5p过表达有效抑制了GC细胞的迁移、克隆形成及增殖(<0.05)。双荧光报告证实miR-191-5p与CDK6的3'UTR结合。WB显示pcDNA-miR-191-5p抑制CDK6表达但促进p21表达。miR-191-5p下调与GC进展相关。miR-191-5p过表达可降低CDK6表达并抑制GC细胞生长。

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