ANKRD13A在肝癌组织中的表达及对其生物学行为的影响  

Expression of ANKRD13A in Hepatocellular Carcinoma and its Effect on Biological Behavior

作　　者：生禹 吴书桢 葛思佳 陈婧 刘肇修[1] 黄伟[1] 陆翠华[1] SHENG Yu;WU Shuzhen;GE Sijia(Department of Gastroenterology,Affiliated Hospital of Nantong University,Jiangsu 226001,China)

机构地区：[1]南通大学附属医院消化内科,226001

出　　处：《医学研究杂志》2023年第9期39-45,共7页Journal of Medical Research

基　　金：国家自然科学基金资助项目(面上项目)(82070624);江苏省研究生科研与实践创新计划项目(SJCX21-1453)。

摘　　要：目的检测锚蛋白重复结构域13A蛋白(ankyrin repeat domain protein 13A)在肝癌患者肝组织中的表达情况,并探究其对肝癌细胞增殖和迁移能力的影响及可能的分子机制。方法利用免疫组织化学、Western blot法以及实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测肝癌患者癌组织和癌旁组织中ANKRD13A的表达水平,并分析其表达水平与患者临床病理参数的相关性;在肝癌细胞系SMMC-7721中,转染目标质粒或空载质粒,通过嘌呤霉素筛选,建立稳定过表达ANKRD13A或空载质粒的细胞系。在体外,分别利用细胞迁移实验、划痕愈合实验以及CCK-8细胞增殖实验,检测过表达ANKRD13A对细胞迁移和增殖能力的影响;在体内,通过裸鼠皮下成瘤实验,进一步验证ANKRD13A对肝癌细胞增殖能力的影响;最后通过信号通路分析,探究ANKRD13A调控肝癌细胞生物学行为可能的分子机制。结果ANKRD13A在肝癌组织中的表达显著低于对应的癌旁组织,并且其表达水平与患者的BCLC分期(P=0.001)和是否合并肝硬化(P=0.003)显著相关。体外细胞模型和体内裸鼠实验结果表明,过表达ANKRD13A可显著抑制肝癌细胞的迁移和增殖能力,减少肝癌细胞中MEK1/2的磷酸化水平。结论ANKRD13A可能通过调控MEK1/2的磷酸化水平进而影响肝癌细胞的增殖和迁移能力,有望成为肝癌治疗的潜在靶点。Objective To examine the expression of ANKRD13A in liver tissue of patients with hepatocellular carcinoma(HCC),and explore the effect of ANKRD13A on the proliferation and migration of HCC cells and its possible molecular mechanism.Methods Immunohistochemistry,Western blot and real-time quantitative polymerase chain reaction(RT-qPCR)were used to detect the expression level of ANKRD13A in cancer tissues and adjacent tissues of HCC patients,and to analyze the correlation between the expression level of ANKRD13A and clinicopathological parameters of the patients.The target plasmid and empty plasmid were transfected into HCC cell lines SMMC-7721,screened by puromycin,then stable overexpression of ANKRD13A or empty plasmid cell lines were established,In vitro,the effect of ANKRD13A on cell migration and proliferation were detected by cell migration assay,scratch healing assay,CCK-8 cell proliferation assay.In vivo,the effect was further verified by subcutaneous tumorigenesis assay in nude mice.Finally,the possible molecular mechanism of ANKRD13A which regulated the biological behavior of HCC cells was also explored.Results The expression of ANKRD13A in HCC tissues was obviously lower than that in adjacent tissues,and its expression level was significantly correlated with BCLC stage(P=0.001)and cirrhosis(P=0.003).The results of in vitro cell model and in vivo nude mouse experiment showed that overexpression of ANKRD13A could significantly inhibit the migration and proliferation of HCC,and reduce the phosphorylation level of MEK1/2 in HCC cells.Conclusion ANKRD13A may influence proliferation and migration of HCC cells by regulating phosphorylation level of MEK1/2,which is expected to become a potential therapeutic target for HCC treatment.

关 键 词：肝细胞癌 细胞增殖 细胞迁移 ANKRD13A 

分 类 号：R735[医药卫生—肿瘤]