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

中国医药导刊 ›› 2021, Vol. 23 ›› Issue (5): 364-370.

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

周志刚1，2，夏红1，刘芳1，苏坚1，3，曾颖1，苏琦1*

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

收稿日期:2021-03-29 修回日期:2021-04-09 出版日期:2021-05-28 发布日期:2021-05-28
基金资助:
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）

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

ZHOU Zhigang1，2， XIA Hong1， LIU Fang1， SU Jian1，3， ZENG Ying1， SU Qi1*

1. 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；
2. Department of Oncology， The First People′s Hospital of Changde City， Hunan Changde 415003， China；
3. Department of Pathology， The Second Affiliated Hospital， University of South China， Hunan Hengyang 421001， China

Received:2021-03-29 Revised:2021-04-09 Online:2021-05-28 Published:2021-05-28
Contact: 苏无琦 E-mail:suqi1945@163.com

摘要/Abstract

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=0.005 4)和空载体组11.59%降低(P=0.005 6)。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<0.000 1)和空载体组（124.1±3.1）mm (P<0.000 1)。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=0.000 3)和空载体组（49.3±3.1）个 (P=0.000 3)。RT-PCR和Western blot显示，LIMK1过表达组LIMK1表达分别高于对照组 (P<0.000 1)和空载体组 (P=0.000 2)，p-LIMK1表达分别高于对照组(P=0.000 3)和空载体组 (P=0.000 4)。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=0.000 9)和空载体组(P=0.001 8)。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。

关键词: font-size:medium, ">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

Abstract: 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=0.005 4) and 11.59% in the empty vector group (P=0.005 6). 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<0.000 1). 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=0.000 3) and （49.3±3.1） cells in the empty vector group (P=0.000 3). RT-PCR and Western blot showed that the LIMK1 expression in the overexpression group was significantly higher than that in the control group (P<0.000 1) and the empty vector group (P=0.000 2)， and the p-LIMK1 expression was significantly higher than that in the control group (P=0.000 3) and the empty vector group (P=0.000 4). 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 higher than that in the control group (P=0.000 9) and the empty vector group (P=0.001 8). Conclusion： The over expression of LIMK1 promote the proliferation， migration and invasion of MGC803 cells by activating LIMK1/Cofilin1 pathway.

Key words: font-size:medium, ">Human gastric cancer MGC803 cells； LIMK1； Eukaryotic expression vector transfection； LIMK1/cofilin1 pathway； Proliferation； Migration； Invasion

中图分类号:

周志刚, 夏红, 刘芳, 苏坚, 曾颖, 苏琦. 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[J]. 中国医药导刊, 2021, 23(5): 364-370.

ZHOU Zhigang, XIA Hong, LIU Fang, SU Jian, ZENG Ying, SU Qi. LIMK1 Overexpression Promotes the Proliferation， Migration and Invasion of Human Gastric Cancer MGC803 Cells through the LIMK1/cofilin1 Pathway[J]. CHINESE JOURNAL OF MEDICINAL GUIDE, 2021, 23(5): 364-370.

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