miR-485-5p调控MEK/ERK信号通路对肝癌细胞增殖、侵袭和凋亡的机制研究  

Mechanism of miR⁃485⁃5p regulating the mek/erk signal pathway on the proliferation,invasion,and apop⁃tosis of hepatocellular carcinoma cells

作　　者：朱艾 刘家芸 李旻珊 李懿 ZHU Ai;LIU Jia-yun;LI Min-shan;LI Yi(920th Hospital of the Joint Logistics Support Force of the People's Liberation Army of China,Kunming,Yunnan Province 650000,China)

机构地区：[1]中国人民解放军联勤保障部队第920医院,云南昆明650000

出　　处：《解剖学研究》2024年第6期566-572,共7页Anatomy Research

基　　金：云南省应用基础研究面上项目(2020FB137)。

摘　　要：目的探讨miR-485-5p调控MEK/ERK信号通路对肝癌细胞增殖、侵袭和凋亡的机制研究。方法选择2021年1月-2022年12月在我院行根治性肝癌切除术的肝癌患者50例,切除肝癌组织及癌旁组织后迅速液氮冷冻,RT-PCR检测组织及细胞中miR-485-5p相对表达量。将肝癌Hep3B细胞随机分为miR-NC组、miR-485-5p组和miR-485-5p+ML099组。CCK-8检测肝癌细胞增殖能力,Transwell法检测细胞侵袭能力,流式细胞仪检测细胞凋亡率,检测细胞中p-MEK、MEK、p-ERK、ERK蛋白表达(蛋白质印迹法)。结果与癌旁组织中miR-485-5p相对表达量(1.18±0.23)相比,肝癌组织(0.19±0.14)明显降低(P<0.01)。与人正常肝细胞株LO2 miR-485-5p相对表达量(1.26±0.23)相比,人肝癌细胞株Huh7(0.36±0.09)、Hep3B(0.17±0.06)和HepG2(0.31±0.07)明显降低(P<0.01)。与miR-NC组相比,miR-485-5p组Hep3B细胞中miR-485-5p相对表达量(1.25±0.18)和细胞凋亡率(18.62%±2.11%)明显升高,细胞活力(24 h:0.23±0.03,48 h:0.39±0.06,72 h:0.57±0.08)、细胞侵袭数目(71.06±8.92)和细胞中p-MEK/MEK比值(0.31±0.04)、p-ERK/ERK比值(0.42±0.05)明显降低(P<0.01);和miR-485-5p组相比,miR-485-5p+ML099组Hep3B细胞中miR-485-5p相对表达量(0.31±0.08)和细胞凋亡率(5.18%±0.64%)明显降低,细胞活力(24 h:0.24±0.04,48 h:0.61±0.08,72 h:0.98±0.10)、细胞侵袭数目(164.11±17.18)和细胞中p-MEK/MEK比值(0.69±0.08)、p-ERK/ERK比值(0.89±0.10)明显升高(P<0.01)。结论miR-485-5p在肝癌中呈低表达,过表达miR-485-5p可能通过抑制MEK/ERK信号通路抑制肝癌细胞增殖和侵袭,并诱导肝癌细胞凋亡。Objective To investigate the mechanism of miR-485-5p regulating the MEK/ERK signaling pathway on the proliferation,invasion,and apoptosis of liver cancer cells.Methods Fifty liver cancer patients who underwent radical resection of liver cancer in our hospital from January 2021 to December 2022 were selected.After resection of liver cancer tissue and adjacent tissues,liquid nitrogen freezing was performed quickly,and the relative expression of miR-485-5p in tissues and cells was detected by RT-PCR.Hep3B cells from liver cancer were randomly divided into miR-NC group,miR-485-5p group,and miR-485-5p+ML099 group.CCK-8 was used to detect the pro-liferation ability of liver cancer cells,Transwell method was used to detect cell invasion ability,flow cytometry was used to detect cell apoptosis rate,and protein blotting was used to detect the expression of p-MEK,MEK,p-ERK,and ERK proteins in the cells.Results Compared with adjacent tissues(1.18±0.23),the relative expression of miR-485-5p in liver cancer tissue was significantly reduced(0.19±0.14)(P<0.01).Compared with the normal human liv-er cell line LO2(1.26±0.23),the relative expression of miR-485-5p in human liver cancer cell lines Huh7(0.36±0.09),Hep3B(0.17±0.06),and HepG2(0.31±0.07)was significantly reduced(P<0.01).Compared with the miR-NC group,the miR-485-5p group showed a significant increase in the relative expression of miR-485-5p(1.25±0.18)and cell apoptosis rate(18.62%±2.11%)in Hep3B cells,while the cell viability(24 h:0.23±0.03,48 h:0.39±0.06,72 h:0.57±0.08),cell invasion number(71.06±8.92),and p-MEK/MEK ratio(0.31±0.04)and p-ERK/ERK ratio(0.42±0.05)were significantly reduced(P<0.01);Compared with the miR-485-5p group,the miR-485-5p+ML099 group showed a significant decrease in miR-485-5p relative expression(0.31±0.08)and apopto-sis rate(5.18±0.64)in Hep3B cells,while cell viability(24 h:0.24±0.04,48 h:0.61±0.08,72 h:0.98±0.10),number of cell invasions(164.11±17.18),and p-MEK/MEK ratio(0.69±0.08)and p-ERK/ERK ratio(0.89±0.10)were signifi

关 键 词：肝癌 微小RNA-485-5p MEK/ERK信号通路 人肝癌细胞株 增殖 迁移 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]