CHMP4C调控坏死性凋亡促进胃癌的作用及机制研究  

Role of CHMP4C in gastric cancer development through regulating necroptosis and its action mechanism

作　　者：郭齐宁 李亚萍 裴莉[2] 于龙臣 罗正东 赵瑞[1] 牛忠芳 张欣 GUO Qi-ning;LI Ya-ping;PEI Li;YU Long-chen;LUO Zheng-dong;ZHAO Rui;NIU Zhong-fang;ZHANG Xin(Department of Clinical Laboratory,Qilu Hospital of Shandong University,Jinan 250012,China;Department 6 of Recuperation,Dalian Rehabilitation Recuperation Center of Joint Logistics Support Force of PLA,Dalian 116013,China)

机构地区：[1]山东大学齐鲁医院检验科,山东济南250012 [2]中国人民解放军联勤保障部队大连康复疗养中心疗养六科,辽宁大连116013

出　　处：《中国现代普通外科进展》2025年第2期125-133,共9页Chinese Journal of Current Advances in General Surgery

基　　金：国家自然科学基金项目(82172339)。

摘　　要：目的:探讨染色质修饰蛋白4C(CHMP4C)调控坏死性凋亡途径在胃癌(GC)发生发展中的作用及机制。方法:通过生物信息学方法分析CHMP4C在泛癌中的表达情况,用RT-qPCR和Western blot检测人正常胃上皮细胞与GC细胞系中CHMP4C的表达情况。在GC细胞系中过表达或敲低CHMP4C,利用CCK-8、平板克隆形成实验检测CHMP4C对GC细胞生长、增殖能力的影响。利用CCK-8实验和Hoechst/PI双染色实验检测经坏死性凋亡诱导剂TSZ或抑制剂necrostatin-1(Nec-1)处理后,GC细胞死亡率和PI阳性细胞比率变化。Western blot检测GC细胞RIPK1、RIPK3、MLKL蛋白质及磷酸化水平。结果:CHMP4C在GC组织和细胞中表达升高。CCK-8和平板克隆形成实验结果显示,过表达CHMP4C的GC细胞增殖能力和集落形成效率明显提高,而敲低CHMP4C的GC细胞明显减弱。并且,CCK-8和Hoechst 33342/PI双染实验结果显示,上调CHMP4C能够抑制TSZ诱导的GC细胞死亡;而Nec-1能够逆转CHMP4C敲低引起的GC细胞活力减低。Western blot结果显示,p-RIPK1、p-RIPK3、p-MLKL水平在过表达细胞中均明显降低,而在敲低细胞中升高;对敲低细胞采用Nec-1处理后,这三种蛋白表达水平下降。结论:CHMP4C可能通过抑制RIPK1/RIPK3/MLKL信号通路的磷酸化负向调控坏死性凋亡促进GC进展,有望成为GC治疗的潜在靶点。Objective:Exploring the role and mechanism of CHMP4C in regulating necroptosis during gastric cancer development and progression.Method:The expression of CHMP4C in pan-cancer was analyzed by bioinformatics methods,and the expression of CHMP4C was detected in human normal gastric epithelial cells and GC cell lines by RTqPCR and Western blot.Overexpression or knockdown of CHMP4C was performed in GC cell lines,and the effects of CHMP4C on the growth and proliferation of GC cells were detected using CCK-8 and clone formation assays.The CCK-8 experiment and Hoechst/PI double staining experiment were used to detect the changes in GC cell mortality and PI positive cell ratio after treatment with the necroptsis inducer TSZ or inhibitor necrostatin-1(Nec-1).Western blot assay was used to detect the protein and phosphorylation levels of RIPK1,RIPK3,and MLKL in GC cells.Result:CHMP4C was upregulated in GC tissues and cells.The CCK-8 and clone formation experiments showed that overexpression of CHMP4C significantly improved the proliferation ability and colony formation efficiency of GC cells,while knockdown of CHMP4C significantly weakened GC cells.Moreover,the results of CCK-8 and Hoechst 33342/PI double staining experiments showed that upregulated CHMP4C could inhibit TSZ induced GC cell death;Nec-1 can reverse the decrease in GC cell viability caused by CHMP4C knockdown.Western blot experiment showed that the levels of p-RIPK1,p-RIPK3,and p-MLKL were significantly decreased in overexpressing cells,while they were increased in knockdown cells.After treatment with Nec-1,the expression levels of these three proteins decreased in knockdown cells.Conclusion:CHMP4C may promote GC progression by negatively regulating necroptosis through inhibiting the phosphorylation of the RIPK1/RIPK3/MLKL signaling pathway,suggesting that it is expected to be a potential target for GC therapy.

关 键 词：胃肿瘤 染色质修饰蛋白4C 坏死性凋亡 靶点 

分 类 号：R735.2[医药卫生—肿瘤]