miR-502-3p抑制结肠癌细胞生长和迁移的作用及机制研究  

Effect and mechanism of miR-502-3p on inhibiting the growth and migration of colorectal cancer cells

作　　者：胡光磊 曹晨曦 王晓光 陈振伟 HU Guanglei;CAO Chenxi;WANG Xiaoguang;CHEN Zhenwei(Graduate School,Bengbu Medical University,Anhui 233030,China;不详)

机构地区：[1]蚌埠医科大学研究生院,233030 [2]嘉兴市第二医院肿瘤外科

出　　处：《浙江医学》2024年第21期2252-2259,I0003,共9页Zhejiang Medical Journal

基　　金：浙江省医药卫生科技计划项目(2021KY354);嘉兴市科技计划项目(2022AD30010)。

摘　　要：目的探讨miR-502-3p抑制结肠癌生长和迁移的相关机制。方法收集2020年11月至2021年12月在嘉兴市第二医院肛肠外科经手术切除且无远处转移的10例结肠癌患者的癌组织及癌旁正常结肠组织,比较miR-502-3p表达水平。取HCT116细胞株构建miR-502-3p模拟物、模拟物对照稳转细胞,观察miR-502-3p对细胞增殖、侵袭、凋亡及生长的影响;取10只裸鼠分别腋下接种所构建的稳转细胞,比较接种miR-502-3p模拟物与模拟物对照裸鼠成瘤情况。采用双荧光素酶报告基因检测miR-502-3p与核转运蛋白α4(KPNA4)的靶向结合关系。观察KPNA4对经miR-502-3p干预后细胞侵袭、凋亡及生长以及KPNA4、N-钙黏蛋白(N-cadherin)、E-钙黏蛋白(E-cadherin)表达的影响,进一步分析KPNA4通过丝裂原活化蛋白激酶(MAPK)信号通路发挥作用的机制。结果结肠癌组织中miR-502-3p表达水平明显低于癌旁正常组织(P=0.028)。相较于模拟物对照组,miR-502-3p模拟物组HCT116细胞株在24、48、72、96 h时增殖能力均明显减弱(均P<0.05),侵袭能力亦明显减弱,细胞凋亡能力增强,且出现细胞周期G1阻滞现象,KPNA4、N-cadherin表达量均明显降低(均P<0.01),E-cadherin表达量明显升高(P<0.01)。相较于模拟物对照,接种miR-502-3p模拟物裸鼠肿瘤生长缓慢,肿瘤体积和重量均有所降低,第23天时比较差异均有统计学意义(均P<0.05)。miR-502-3p模拟物能明显降低野生型KPNA4的荧光素酶活性(P<0.001),但对突变型KPNA4的荧光素酶活性无明显影响(P=0.158)。联合KPNA4后,miR-502-3p干预后的HCT116细胞株侵袭和生长能力均增强,凋亡能力下降,KPNA4、N-cadherin表达量均明显升高(均P<0.01),E-cadherin表达量无明显变化(P>0.05)。相较于KPNA4过表达组,KPNA4过表达+MAPK抑制剂组在24、48、72、96 h的细胞增殖能力均明显降低(均P<0.05),细胞侵袭和生长能力均明显减弱,细胞凋亡能力增强。结论miR-502-3p可抑制结肠癌细胞�Objective To explore the underlying mechanisms by which miR-502-3p inhibits the growth and migration of colorectal cancer.Methods Cancer tissues and adjacent normal tissues were collected from 10 colorectal cancer patients who underwent surgical resection without distant metastasis at the Second Hospital of Jiaxing City from November 2020 to December 2021.The expression levels of miR-502-3p were compared.HCT116 cell line was used to construct miR 502-3p mimic and control stable-transfected cells.The effects of miR-502-3p on cell proliferation,invasion,apoptosis,and growth were observed.Ten nude mice were inoculated with the constructed stable-transfected cells under the armpit,and the tumorigenicity was compared between mice injected with miR 502-3p mimic and control mice.The targeted binding relationship between miR 502-3p and karyopherin-alpha 4(KPNA4)was detected using a dual-luciferase reporter gene assay.The effects of KPNA4 on cell invasion,apoptosis,and growth after miR 502-3p intervention,as well as the expressions of KPNA4,N-cadherin,and E-cadherin,were observed.Furthermore,the mechanism of KPNA4's action through the mitogen-activated protein kinase(MAPK)signaling pathway was analyzed.Results The expression level of miR-502-3p in colorectal cancer tissues was significantly lower than that in adjacent normal tissues(P=0.028).Compared with the control group,the proliferative capacity of HCT116 cells transfected with miR-502-3p mimic was significantly weakened at 24,48,72,and 96 hours(all P<0.05).The invasive ability was also significantly weakened,and the apoptosis ability was enhanced,accompanied by G1 phase arrest.The expression levels of KPNA4 and N-cadherin were significantly decreased(both P<0.01),while the expression level of E-cadherin was significantly increased(P<0.01).Compared with the control group,the mice injected with miR 502-3p mimic showed slower tumor growth,reduced tumor volume and weight,and significant differences were observed on the 23rd day(all P<0.05).miR 502-3p mimic significantly

关 键 词：结肠癌 miRNA-502-3p 核转运蛋白α4 上皮-间质转化 丝裂原活化蛋白激酶 

分 类 号：R735.35[医药卫生—肿瘤]