李英,王兴,李永宁,刘鹏,刘松柏,潘耀振
LI Ying,WANG Xing,LI Yongning,LIU Peng,LIU Songbai,PAN Yaozhen

• 1:贵州医科大学附属肿瘤医院肝胆外科

摘要(Abstract)：

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(CoIP)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.

关键词(KeyWords)： 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
hepatocellular carcinoma;F-box and WD repeat domain-containing protein 7;minichromosome maintenance complex component 7;cell proliferation;MHCC-97H cell

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81960535);; 贵州省科技计划项目(黔科合平台人才-﹝2018﹞5779-29);; 贵阳市科技计划项目(筑科合同﹝2018﹞1-86号)

作者(Author): 李英,王兴,李永宁,刘鹏,刘松柏,潘耀振
LI Ying,WANG Xing,LI Yongning,LIU Peng,LIU Songbai,PAN Yaozhen

DOI: 10.19367/j.cnki.2096-8388.2024.02.005

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