CHMP4C通过调控PI3K/AKT信号通路影响NSCLC细胞增殖与凋亡  

CHMP4C affects NSCLC cell proliferation and apoptosis by regulating the PI3K/AKT signaling pathway

作　　者：任碧 苟浩铖 张琴[1,2] 贺丽萍[1,2] 薛林峰 孙金洪 蒋莉 REN Bi;GOU Haocheng;ZHANG Qin;HE Liping;XUE Linfeng;SUN Jinhong;JIANG Li(Department of Respiratory and Critical Care Medicine,Affiliated Hospital of North Sichuan Medical College,Sichuan Nanchong 637000,China;North Sichuan Medical College,Sichuan Nanchong 637000,China;Department of Otolaryngology,Head and Neck Surgery,Nanchong Central Hospital,the Second Clinical Medical College of North Sichuan Medical College,Sichuan Nanchong 637000,China)

机构地区：[1]川北医学院附属医院呼吸与危重症医学科,四川南充637000 [2]川北医学院,四川南充637000 [3]南充市中心医院耳鼻咽喉头颈外科,川北医学院第二临床医学院,四川南充637000

出　　处：《现代肿瘤医学》2024年第9期1580-1588,共9页Journal of Modern Oncology

基　　金：四川省科技计划项目(编号:2021YFS0572);四川省医学会项目(编号:2021HR24)。

摘　　要：目的:探究染色质修饰蛋白4C(CHMP4C)对非小细胞肺癌(NSCLC)进展及顺铂敏感性的影响及其机制。方法:通过免疫组化检测CHMP4C在NSCLC癌组织和癌旁组织中的表达情况。利用实时荧光定量PCR(RT-qPCR)和蛋白免疫印迹法(Western blot)检测CHMP4C在NSCLC细胞系中的表达水平。细胞计数试剂盒(CCK-8)和细胞克隆实验检测细胞活力及半数抑制浓度IC50;流式细胞术检测细胞周期和凋亡。Western blot分析细胞凋亡相关蛋白(caspase 3、Bad)及PI3K/AKT信号通路中关键蛋白的表达水平。另外,我们采用动物模型进一步验证CHMP4C对体内肿瘤生长的影响。结果:CHMP4C在NSCLC癌组织和细胞系中高表达,与肿瘤T分期显著相关(P<0.05)。敲低CHMP4C抑制细胞增殖,促进细胞凋亡,并将细胞阻滞在S期(P<0.05)。敲低CHMP4C可抑制PI3K/AKT信号通路活化,然而激活PI3K/AKT信号通路逆转了CHMP4C敲低对NSCLC细胞的影响(P<0.05)。利用顺铂处理细胞后发现NSCLC细胞生长受抑制,且CHMP4C表达降低;敲低CHMP4C联合顺铂治疗明显诱导肿瘤细胞凋亡,抑制肿瘤细胞增殖(P<0.05)。在动物实验中,CHMP4C敲低的肿瘤体积小于对照组,其ki67表达显著降低,而caspase 3表达升高(P<0.05)。结论:CHMP4C在NSCLC癌组织及细胞系中高表达,在体内及体外实验中敲低CHMP4C可抑制NSCLC细胞增殖,促进细胞凋亡,增强顺铂治疗的敏感性;CHMP4C可能通过PI3K/AKT信号通路参与NSCLC进展与顺铂耐药。Objective:To investigate the effect of chromatin modifying protein 4C(CHMP4C)on the progression and cisplatin sensitivity of non-small cell lung cancer(NSCLC)and its underlying mechanism.Methods:Immunohistochemistry(IHC)was used to analyze the expression level of CHMP4C in NSCLC tumor tissues and paracancerous tissues.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blot were used to detect the expression levels of CHMP4C in NSCLC cell lines.Moreover,CCK-8 and cell clone assays were used to detect cell proliferation and IC_(50)(half inhibitory concentration,IC_(50)).Cell cycle and apoptosis were analyzed by flow cytometry.Western blot analyses were performed to examine the expression of caspase 3,Bad and the key proteins in the PI3K/AKT pathway.In addition,we used an animal model to further validate the effect of CHMP4C on tumor growth in vivo.Results:CHMP4C was highly expressed in NSCLC tissues and cell lines,and correlated with T stage(P<0.05).Knockdown of CHMP4C inhibited cell proliferation and promoted apoptosis by blocking the S phase of cell cycle(P<0.05).Moreover,it was found that knockdown of CHMP4C inhibited the activation of the PI3K/AKT signaling pathway,however it's activator reversed the effect of CHMP4C knockdown on NSCLC cells(P<0.05).Additionally,treatment of NSCLC cells with cisplatin revealed that cell growth was inhibited and CHMP4C expression was reduced.Cisplatin treatment of CHMP4C knockdown cells significantly induced tumor cell apoptosis and inhibited cell proliferation(P<0.05).In animal experiments,the tumor volume of CHMP4C knockdown group was smaller than the control group,and its ki67 expression was significantly reduced while caspase 3 expression was increased(P<0.05).Conclusion:The expression of CHMP4C in NSCLC tumor tissues and cell lines was increased.Knockdown of CHMP4C in vivo and in vitro inhibited NSCLC cell proliferation,promoted apoptosis,and enhanced the sensitivity of cisplatin to cells.CHMP4C may be involved in NSCLC progression and cisplatin resistance through

关 键 词：CHMP4C 非小细胞肺癌 PI3K/AKT信号通路 细胞增殖 细胞凋亡 顺铂 

分 类 号：R734.2[医药卫生—肿瘤]