ANP32A对结肠癌细胞增殖的影响及机制研究  

Effect of ANP32A on the proliferation of colon cancer cells and related mechanism

作　　者：徐光齐 荀江[1] 芮鸿庆 濮阳永强 周晓华[1] XU Guangqi;XUN Jiang;RUI Hongqing;PUYANG Yongqiang;ZHOU Xiaohua(Department of General Surgery,Gaochun People′s Hospital,Nanjing,Jiangsu 211300,China)

机构地区：[1]高淳人民医院普通外科,江苏南京211300

出　　处：《徐州医科大学学报》2023年第11期847-854,共8页Journal of Xuzhou Medical University

基　　金：江苏卫生健康学院面上项目(JKC2021056)。

摘　　要：目的探讨酸性核磷蛋白32A(ANP32A)对结肠癌细胞增殖的影响及相关机制。方法利用生物信息学分析结肠癌组织中ANP32A mRNA水平及其与患者预后的关系。在DLD-1和HCT-116细胞中构建稳定沉默ANP32A的慢病毒细胞系,并用CCK-8和集落形成实验检测细胞增殖能力。采用瞬时转染的方法将myc-ANP32A质粒转入DLD-1和HCT-116细胞,并用CCK-8和集落形成实验检测细胞增殖能力。利用实时荧光定量PCR(qPCR)和Western blot检测ANP32A对高迁移率族蛋白B1(HMGB1)表达的影响。利用Western blot检测ANP32A对细胞外调节蛋白激酶(ERK)通路的影响。最后在沉默ANP32A的细胞中转入flag-HMGB1质粒,并利用Western blot、CCK-8检测ERK通路的蛋白水平和结肠癌细胞的增殖能力。结果结肠癌组织中ANP32A mRNA高表达,ANP32A mRNA高表达的结肠癌患者预后较差。沉默ANP32A可以减弱结肠癌细胞的增殖能力,过表达ANP32A可以增强结肠癌细胞增殖能力。ANP32A可以增强HMGB1的转录进而促进其蛋白表达。ANP32A可以通过HMGB1激活ERK信号通路进而促进结肠癌细胞增殖。结论ANP32A在结肠癌中高表达,通过调控HMGB1转录增强其蛋白表达,并激活ERK通路促进结肠癌细胞增殖。Objective To investigate the effect of acidic nuclear phosphoprotein 32A(ANP32A)on the proliferation of colon cancer cells and related mechanisms.Methods The mRNA level of ANP32A in colon cancer and its relationship with patient survival were analyzed by bioinformatics.DLD-1 and HCT-116 cells were used to construct stable lentiviral cell lines with ANP32A silencing,and cell proliferation was detected by CCK-8 and colony formation assays.Myc-ANP32A plasmid was transferred into DLD-1 and HCT-116 cells by transient transfection,and cell proliferation was detected by CCK-8 and colony formation assays.The effect of ANP32A on the expression of high mobility group protein B1(HMGB1)was detected by qPCR and Western blot.The effect of ANP32A on extracellular regulated protein kinase(ERK)pathway was detected by Western blot.Finally,flag-HMGB1 plasmid was transferred into ANP32A silenced cells,and the levels of the proteins in the ERK pathway and the proliferation ability of colon cancer cells were detected by Western blot and CCK-8.Results The mRNA levels of ANP32A in colon cancer were high and negatively correlated with survival.ANP32A silencing attenuated the proliferation of colon cancer cells,while ANP32A overexpression enhanced the proliferation of the cells.ANP32A enhanced the transcription of HMGB1 and promoted its protein expression.ANP32A promoted the proliferation of colon cancer cells by activating the ERK signaling pathway via HMGB1.Conclusions ANP32A is highly expressed in colon cancer.It can promote HMGB1 protein expression by enhancing its transcription,thereby activates the ERK pathway to promote colon cancer cell proliferation.

关 键 词：结肠癌 酸性核磷蛋白32A 高迁移率族蛋白B1 细胞增殖 蛋白激酶 

分 类 号：R735.3[医药卫生—肿瘤]