首页|FBXW7/MCM7信号轴介导肝癌细胞增殖的研究

FBXW7/MCM7信号轴介导肝癌细胞增殖的研究

FBXW7/MCM7 signaling pathway involved in proliferation of hepatocellular carcinoma cells

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目的 探讨FBXW7靶向调控MCM7表达对肝癌细胞增殖的影响.方法 采用免疫共沉淀法(Co-IP)和免疫荧光染色法(IF),分析肝癌细胞系MHCC-97H中FBXW7与MCM7的互作关系;通过慢病毒感染过表达FBXW7和(或)MCM7,小干扰RNA技术下调FBXW7和(或)MCM7的表达;采用Western blot检测FBXW7对MCM7蛋白表达的影响,CCK-8实验检测肝癌细胞的增殖活性,EdU细胞增殖检测分析肝癌细胞的增殖能力,平板克隆实验分析肝癌细胞的集落形成能力.结果 在MHCC-97H细胞中,FBXW7与MCM7存在互作关系:过表达FBXW7抑制了 MCM7的蛋白表达水平、并抑制了肝癌MHCC-97H细胞的增殖(P＜0.05),在过表达FBXW7的基础上增加MCM7的表达后,细胞增殖能力提高(P＜0.05);下调FBXW7后细胞中MCM7表达增加、细胞增殖能力增强(P＜0.05),而下调FBXW7的同时干扰MCM7表达后,细胞增殖能力减弱(P＜0.05).结论 FBXW7能抑制肝癌细胞增殖,其机制与负向调节肝癌细胞中促癌蛋白MCM7的表达水平有关.
Objective To explore the effect of FBXW7 targeted regulation of MCM7 expression on the proliferation of hepatocellular carcinoma(HCC).Methods The interaction between FBXW7 and MCM7 in MHCC-97H hepatoma cells was detected by co-immunoprecipitation(Co-IP)and immunofluorescence staining(IF).The expression of FBXW7 and/or MCM7 in MHCC-97H cells was overexpressed by lentiviral infection,and down-regulated by small interfering RNA technique.The effect of FBXW7 on the expression of MCM7 protein in MHCC-97H cells was detected by Western blot.CCK-8 and EdU assays were adopted to test the proliferation activity of MHCC-97H cells.The colony formation ability of hepatoma cells was analyzed by plate cloning experiment.Results There is an interaction between FBXW7 and MCM7 in MHCC-97H cells.Overexpression of FBXW7 inhibited the expression of MCM7 and blocked the proliferation of hepatoma cells(P＜0.05).The ability of cell proliferation was improved when MCM7 increased based on the overexpression of FBXW7(P＜0.05).After down-regulation of FBXW7,the expression of MCM7 increased and the ability of cell proliferation was enhanced(P＜0.05).However,after down-regulation of FBXW7 and interference of the expression of MCM7,the cell proliferation decreased(P＜0.05).Conclusion FBXW7 can inhibit the proliferation of HCC cells by negatively regulating the expression of oncoprotein MCM7 in hepatoma cells.

hepatocellular carcinomaF-box and WD repeat domain-containing protein 7minichromosome maintenance complex component 7cell proliferationMHCC-97H cell

李英、王兴、李永宁、刘鹏、刘松柏、潘耀振

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贵州医科大学附属肿瘤医院肝胆外科,贵州贵阳 550000

肝癌 F-box和WD重复结构域蛋白7 微小染色体维持蛋白-7 细胞增殖 MHCC-97H细胞

国家自然科学基金贵州省科技计划项目贵阳市科技计划项目

81960535黔科合平台人才-[2018]5779-29筑科合同[2018]1-86号

2024

10.19367/j.cnki.2096-8388.2024.02.005

贵州医科大学学报
贵阳医学院

贵州医科大学学报

CSTPCD
影响因子：0.827
ISSN：2096-8388
年,卷(期)：2024.49(2)
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