MYPT1通过RhoAROCK信号通路抑制结直肠癌细胞增殖和迁移

MYPT1 inhibits colorectal cancer cell growth and migration by RhoA/ROCK signaling pathway

目的 探讨实肌球蛋白磷酸酶靶亚基1(MYPT1)在结直肠癌发生发展中的作用及其可能的作用机制。方法 RT-PCR检测mRNA表达水平,Western blot检测蛋白表达水平,CCK-8法检测细胞增殖活性,Transwell法检测结直肠癌细胞迁移能力,生物信息学软件筛选可与MYPT1相互作用的蛋白质,通过TCGA数据库分析MYPT1表达水平及与RAS同源基因家族成员A(RhoA)和锌离子相关的RhoA蛋白1(ROCK1)的相关性。根据实验方案进行分组,分为空白对照组、阴性对照组、MYPT1过表达组、RhoA过表达组、MYPT1过表达+RhoA空载组和MYPT1过表达+RhoA过表达组。结果 和癌旁正常组织及正常肠粘膜细胞相比,MYPT1在结直肠癌组织和细胞中低表达(P<0.05)。过表达MYPT1可抑制结直肠癌细胞增殖和迁移(P<0.05)。MYPT1负调控RhoA和ROCK1的表达(P<0.05)。过表达RhoA促进结直肠癌细胞增殖和迁移(P<0.05)。MYPT1通过下调RhoA的表达抑制ROCK1表达(P<0.05)。MYPT1通过负调控RhoA抑制细胞增殖和迁移(P<0.05)。结论 MYPT1在结直肠癌组织和细胞中低表达,其通过RhoA/ROCK信号通路调控结直肠癌细胞增殖和迁移。

Objective This study aimed to demonstrate the role of MYPT1(Myosin phosphatase-targeting subunit 1)in the development of colorectal cancer(CRC)and determine the potential mechanisms of action.Methods MRNA expression level was detected by RT-PCR,protein expression level was detected by Western blot,cell proliferation activity was detected by CCK-8 method,migration ability of colorectal cancer cells was detected by Transwell method,protein that could interact with MYPT1 was screened by bioinformatics software.The expression level of MYPT1 and its correlation with RhoA protein 1(ROCK1),A member of RAS homologous gene family,were analyzed by TCGA database.According to the experimental plan,groups were divided into blank control group,negative control group,MYPT1 overexpression group,RhoA overexpression group,MYPT1 overexpression+RhoA no-load group and MYPT1 overexpression+RhoA overexpression group.Results Compared with normal adjacent tissues and normal intestinal mucosal cells,MYPT1 was lower expressed in tissues and cells of colorectal cancer(P<0.05).Overexpression of MYPT1 inhibited the proliferation and migration of colorectal cancer cells(P<0.05).MYPT1 negatively regulates the expression of RhoA and ROCK1(P<0.05).Overexpression of RhoA promotes the proliferation and migration of colorectal cancer cells(P<0.05).This paper pressume that MYPT1 inhibited ROCK1 expression by downregulating RhoA expression(P<0.05).MYPT1 inhibited cell proliferation and migration by negatively regulating RhoA(P<0.05).Conclusion MYPT1 is lowly expressed in CRC cancer tissue and cells,and MYPT1 regulates the proliferative and migratory abilities via the RhoA/ROCK pathway.