LIMK1过表达通过LIMK1/cofilin1通路促进人胃癌MGC803细胞增殖与迁移侵袭  

LIMK1 Overexpression Promotes the Proliferation,Migration and Invasion of Human Gastric Cancer MGC803 Cells through the LIMK1/cofilin1 Pathway

作　　者：周志刚[1,2] 夏红 刘芳[1] 苏坚[1,3] 曾颖[1] 苏琦 ZHOU Zhigang;XIA Hong;LIU Fang;SU Jian;ZENG Ying;SU Qi(Cancer Research Institute,Center for Gastric Cancer Research of Hunan Province,Hunan Province Key Laboratory of Cancer Cellular and Molecular Pathology,University of South China,Hunan Hengyang 421001,China;Department of Oncology,The First People′s Hospital of Changde City,Hunan Changde 415003,China;Department of Pathology,The Second Affiliated Hospital,University of South China,Hunan Hengyang 421001,China)

机构地区：[1]湖南省肿瘤细胞与分子病理学重点实验室,湖南省胃癌研究中心,南华大学肿瘤研究所,湖南衡阳421001 [2]常德市第一人民医院肿瘤科,湖南常德415003 [3]南华大学附属第二医院病理科,湖南衡阳421001

出　　处：《中国医药导刊》2021年第5期364-370,共7页Chinese Journal of Medicinal Guide

基　　金：国家自然科学基金(项目编号:81973532;项目名称:DADS下调DJ-1负调控PTEN/Akt通路抑制人胃癌细胞EMT与侵袭和抗药性);国家自然科学基金(项目编号:81374013;项目名称:二烯丙基二硫上调RORα拮抗Wnt/β-catenin通路抑制人胃癌细胞EMT与迁移侵袭)。

摘　　要：目的:探讨LIMK1过表达对人胃癌MGC803细胞增殖与迁移侵袭的影响。方法:真核表达载体转染技术构建过表达LIMK1基因MGC803细胞;RT-PCR和Western blot检测LIMK1、Cofilin1、p-Cofilin1表达;MTT、流式细胞术、细胞划痕和侵袭实验分别检测LIMK1过表达对MGC803细胞增殖、细胞周期、迁移与侵袭能力的影响。结果:成功构建稳定过表达LIMK1基因MGC803细胞。MTT显示,LIMK1过表达组细胞的增殖活性呈时间依赖性,高于对照组和空载体组(P<0.001)。流式细胞术检测显示,LIMK1过表达组G2/M期细胞百分率8.36%,较MGC803组11.45%(P=0.0054)和空载体组11.59%降低(P=0.0056)。划痕实验显示,24 h后LIMK1过表达组迁移距离(163.9±1.5)mm分别高于MGC803组(127.1±2.0)mm(P<0.0001)和空载体组(124.1±3.1)mm(P<0.0001)。侵袭实验显示,LIMK1过表达组穿膜细胞(86.3±4.2)个分别明显多于MGC803组(48.3±2.5)个(P=0.0003)和空载体组(49.3±3.1)个(P=0.0003)。RT-PCR和Western blot显示,LIMK1过表达组LIMK1表达分别高于对照组(P<0.0001)和空载体组(P=0.0002),p-LIMK1表达分别高于对照组(P=0.0003)和空载体组(P=0.0004)。LIMK1过表达组Cofilin1 mRNA与蛋白表达较MGC803组和空载体组差异均无统计学意义(P>0.05),而p-Cofilin1表达分别高于对照组(P=0.0009)和空载体组(P=0.0018)。结论:LIMK1过表达可通过激活LIMK1/cofilin1通路促进MGC803细胞增殖与迁移侵袭。Objective:To investigate the effect of LIMK1 overexpression on proliferation,migration and invasion of human gastric cancer MGC803 cells.Methods:Eukaryotic expression vector transfection was used to construct MGC803 cells over expressing LIMK1 gene.The expressions of LIMK1,Cofilin1 and p-Cofilin1 were detected by RT-PCR and Western blot.MTT,flow cytometry,cell scratch and invasion assay were used to detect the effects of LIMK1 over expression on the proliferation,cell cycle,migration and invasion of MGC803 cells.Results:MGC803 cells over expressing LIMK1 gene were successfully constructed.MTT showed that the proliferative activity of the cells in the LIMK1 overexpression group,which was in a time-dependent manner,was significantly higher than that in the control group and the empty vector group(P<0.001).Flow cytometry showed that the percentage of G2/M phase cells in LIMK1 overexpression group was 8.36%,which was lower than 11.45%in the MGC803 group(P=0.0054)and 11.59%in the empty vector group(P=0.0056).The scratch test showed that the migration distance after 24 h of the LIMK1 overexpression group was(163.9±1.5)mm which was significantly higher than(127.1±2.0)mm of the MGC803 group and(124.1±3.1)mm of the empty vector group(P<0.0001).Invasion experiments showed that there were(86.3±4.2)transcembrane cells in the LIMK1 overexpression group,which were significantly higher than(48.3±2.5)cells in the MGC803 group(P=0.0003)and(49.3±3.1)cells in the empty vector group(P=0.0003).RT-PCR and Western blot showed that the LIMK1 expression in the overexpression group was significantly higher than that in the control group(P<0.0001)and the empty vector group(P=0.0002),and the p-LIMK1 expression was significantly higher than that in the control group(P=0.0003)and the empty vector group(P=0.0004).The Cofilin1 mRNA and protein expression of the LIMK1 overexpression group was not significantly different from that in the MGC803 group and the empty vector group(P>0.05),while the expression of p-Cofilin1 was significantly hig

关 键 词：人胃癌MGC803细胞 LIMK1 真核表达载体转染 LIMK1/cofilin1通路 增殖 迁移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]