微小RNA-642a-5p通过靶向作用于Ⅰ型胶原α1对结肠癌细胞的迁移和侵袭的影响  

Effect of microRNA-642a-5p on the migration and invasion of colon cancer cells by targeting typeⅠcollagenα1

作　　者：郑光威[1] 方志鹏[1] 林劲权[1] 林振吕[1] 郑建涛[2] Zheng Guangwei;Fang Zhipeng;Lin Jinquan;Lin Zhenlyn;Zheng Jiantao(Department of Emergency Surgery,the First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China;Department of Gastrointestinal Surgery,the First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China)

机构地区：[1]福建医科大学附属第一医院急诊外科,350005 [2]福建医科大学附属第一医院胃肠外科,350005

出　　处：《中华实验外科杂志》2021年第8期1511-1514,共4页Chinese Journal of Experimental Surgery

基　　金：福建省卫生健康中青年骨干人才培养项目(2020GGB030);福建省自然科学基金面上项目(2020J01977)。

摘　　要：目的探讨微小RNA-642a-5p(miR-642a-5p)对结肠癌细胞株SW620细胞迁移和侵袭的影响及分子机制。方法将人结肠癌细胞SW620细胞分成4个组:对照组(OE-NC+mimic-NC组:共转染miR-642a-5p mimics阴性对照与空载体);过表达miR-642a-5p组(OE-NC+mimic-miR-642a-5p组:共转染miR-642a-5p mimics与空载体);过表达miR-642a-5p与Ⅰ型胶原α1(COL1A1)组(OE-COL1A1+mimic-miR-642a-5p组:共转染miR-642a-5p mimics与COL1A1过表达载体);过表达COL1A1组(OE-COL1A1+mimic-NC组:共转染miR-642a-5p mimics阴性对照与COL1A1过表达载体)。反转录-聚合酶链反应(RT-PCR)检测各组细胞内miR-642a-5p及COL1A1的表达;划痕实验、Transwell小室实验检测细胞迁移和侵袭能力;蛋白质印迹法(Western blot)实验检测细胞内COL1A1蛋白表达。两组间比较采用独立样本t检验。结果RT-PCR结果显示应用miR-642a-5p mimics转染细胞后,miR-642a-5p表达水平显著增加;双荧光素酶报告基因实验结果miR-642a-5p直接靶向作用于COL1A1。应用(COL1A1 overexpression,OE-COL1A1)转染后,细胞COL1A1表达水平明显增加。而OE-NC+mimic-miR-642a-5p组和OE-COL1A1+mimic-miR-642a-5p组的RNA相对表达量(18.62±2.31),(2.52±0.44)明显高于OE-NC+mimic-NC组(0.52±0.11),差异有统计学意义(t=36.120、6.125,P<0.05),且OE-COL1A1+mimic-miR-642a-5p组的COL1A1相对表达量(5.88±1.06)明显高于OE-NC+mimic-miR-642a-5p组(0.22±0.04),差异有统计学意义(t=10.201,P<0.05)。与OE-NC+mimic-NC组比较,miR-642a-5p的过表达显著抑制了细胞的迁移和侵袭能力,而过表达COL1A1显著促进SW620细胞的迁移和侵袭能力,并部分逆转miR-642a-5p对细胞迁移和侵袭的抑制作用。结论miR-642a-5p可通过调节COL1A1的表达水平来抑制结肠癌细胞的迁移与侵袭。Objective To investigate the effect of microRNA-642a-5p(miR-642a-5p)on the migration and invasion of colon cancer cell line SW620 and its molecular mechanism.Methods The SW620 cells were divided into 4 groups:control group;miR-642a-5p overexpression group;miR-642a-5p overexpression+collagen type Ⅰ α1(COL1A1)group;COL1A1 overexpression group.Reverse transcriptase-polymerase chain reaction(RT-PCR)was used to detect the expression of miR-642a-5p and COL1A1 in each group.The scratch test and the Transwell chamber test were used to detect cell migration and invasion ability.Western blotting was used to detect the expression of COL1A1 protein.Comparison between two groups was done with independent sample t-test.Results RT-PCR results showed that after transfecting cells with miR-642a-5p mimics,the expression level of miR-642a-5p increased significantly.The results of dual luciferase reporter gene experiments showed that miR-642a-5p directly targeted COL1A1.After transfection with COL1A1 overexpression,the expression level of COL1A1 in cells increased significantly.The RNA and protein expression levels in miR-642a-5p overexpression group and COL1A1+miR-642a-5p group(18.62±2.31,2.52±0.44)were significantly higher than those in control group(0.52±0.11,t=36.120,6.125,P<0.05),and the COL1A1 expression level in the COL1A1+miR-642a-5p group(5.88±1.06)was significantly higher than that in the miR-642a-5p overexpression group(0.22±0.04,t=10.201,P<0.05).Compared with the control group,the overexpression of miR-642a-5p significantly inhibited the migration and invasion ability of cells,while the overexpression of COL1A1 significantly promoted,or partially reverved the migration and invasion ability of SW620 cells.MiR-642a-5p could inhibit cell migration and invasion.Conclusion miR-642a-5p can inhibit the migration and invasion of colon cancer cells by regulating the expression level of COL1A1.

关 键 词：微小RNA 结肠癌 迁移 侵袭 

分 类 号：R735.35[医药卫生—肿瘤]