适应素耳结合外壳相关蛋白2高表达对肝细胞癌生物学功能的影响  

Effect of high expression of adaptin-ear-binding coat-associated protein 2 on the biological function of hepatocellular carcinoma

作　　者：刘杰[1,2] 彭真 程诺 曾志生 李赛嘉 袁文豪 陈二宝 林泽伟 Liu Jie;Peng Zhen;Cheng Nuo;Zeng Zhisheng;Li Saijia;Yuan Wenhao;Chen Erbao;Lin Zewei(Hubei University of Medicine,Shiyan 442000,China;Department of Hepatobiliary Surgery,Peking University Shenzhen Hospital,Shenzhen 518036,China)

机构地区：[1]湖北医药学院,十堰442000 [2]北京大学深圳医院肝胆胰外科,深圳518036

出　　处：《中华实验外科杂志》2024年第5期959-962,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年项目(82303446);深圳市科技计划项目(JCYJ20190809095801653、JCYJ20190809100217290)。

摘　　要：目的:探讨适应素耳结合外壳相关蛋白2(NECAP2)在肝细胞癌中的表达与肝细胞癌(HCC)患者预后的关系,观察NECAP2在肝癌细胞MHCC-97H中的表达及其对HCC增殖及侵袭的影响。方法:从肿瘤基因组图谱(TCGA)数据库下载并整理TCGA-LIHC的数据进行分析;选取16例肝癌样本及其配对的癌旁样本进行研究,用荧光实时定量聚合酶链反应(RT-qPCR)检测这16对样本、正常肝细胞LO2以及肝癌细胞MHCC-97H中的差异表达水平;用小干扰RNA转染MHCC-97H细胞以沉默NECAP2的表达,实验分为实验组(si-NECAP2组)和对照组(si-NC组)。并采用细胞计数试剂盒(CCK-8)、5-乙炔基-2’脱氧尿嘧啶核苷(EdU)实验、Transwell实验和蛋白质印迹法(Western blot)分别检测实验组和对照组的细胞活力、增殖、迁移和侵袭能力以及表皮生长因子受体(EGFR)蛋白的表达量。两组间比较采用t检验。结果:基于TCGA数据库中的肝癌细胞组和正常肝细胞组,癌旁组织中NECAP2的表达水平明显低于肝癌组织(3.747±0.474比4.367±0.593),差异有统计学意义(t=7.102,P<0.05);NECAP2在正常肝组织中的表达(3.746±0.474比4.263±0.503)要显著低于配对的肝癌组织,差异有统计学意义(t=5.152,P<0.05)。ROC曲线下面积(AUC)值为0.802,表明NECAP2对HCC有很好的诊断价值,Kaplan-Meier分析结果显示,NECAP2高表达组患者的生存期更短;实验组细胞吸光度(A)值、EdU阳性细胞占比、迁移细胞数量和侵袭细胞数量[3.201±0.300、(15.06±8.90)%、(103.000±7.550)个、(112.333±5.508)个]明显低于对照组[4.852±0.462、(49.070±10.520)%、(141.000±8.888)个、(146.333±4.041)个],差异有统计学意义(t=5.995、4.269、5.664、8.621,P<0.05)。此外,EGFR的表达量更低,与NECAP2的表达呈正相关。结论:NECAP2在HCC中的表达显著上调,NECAP2高表达组的患者预后更差;细胞实验证明下调NECAP2的表达可能通过下调EGFR的表达来抑制HCC的增殖、迁移及侵袭能力。Objective To investigate the relationship between the expression of adaptin-ear-binding coat-associated protein 2(NECAP2)in hepatocellular carcinoma(HCC)and the prognosis of HCC patients,and to study the expression of NECAP2 in MHCC-97H and its effect on HCC proliferation and invasion.Methods The cancer genome atlas(TCGA)-LIHC were downloaded from TCGA database.A total of 16 HCC samples and their paired adjacent samples were selected and the differential expression levels of normal liver cells and MHCC-97H were detected by real-time fluorescent quantitative polymerase chain reaction(RT-qPCR).MHCC-97H cells were transfected with small interfering RNA to silence NECAP2 expression,and si-NECAP2 group and si-NC group were set up.Cell viability,proliferation,migration,invasion,and epidermal growth factor receptor(EGFR)protein expression were determined by cell counting kit(CCK-8),5-ethynyl-2′-deoxyuridine(EdU),Transwell,and Western blotting.Comparison between the two groups was performed by using the t-test.Results Based on the HCC cell group and normal hepatocyte group in TCGA database,the expression level of NECAP2 in adjacent tissues was significantly lower than that in HCC tissues(3.747±0.474 vs.4.367±0.593,t=7.102,P<0.05);The NECAP2 expression in normal liver tissues was significantly lower than that in paired HCC tissues(3.746±0.474 vs.4.263±0.503,t=5.152,P<0.05).Area under the ROC curve(AUC)value of 0.802 showed that NECAP2 had a good diagnostic value for HCC.The results of the Kaplan-Meier analysis showed that NECAP2 patients in the high expression group had a shorter survival period.The absorbance values,proportion of EdU positive cells,number of migrating cells and number of invasive cells[3.201±0.300,(15.06±8.90)%,(103.000±7.550)cells,(112.333±5.508)cells]were significantly reduced in si-NECAP2 group as compared with those in si-NC group[(4.852±0.462),(49.070±10.520)%,(141.000±8.888)cells,(146.333±4.041)cells,t=5.995,4.269,5.664,8.621,P<0.05].Moreover,EGFR expression was even lower and positiv

关 键 词：适应素耳结合外壳相关蛋白2 肝细胞癌 预后 增殖 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]