内质网跨膜蛋白EVA1A对肝细胞癌Huh7细胞生物学行为的影响  

EFFECT OF ENDOPLASMIC RETICULUM TRANSMEMBRANE PROTEIN EVA1A ON THE BIOLOGICAL BEHAVIOR OF HEPATOCELLULAR CARCINOMA Huh7 CELLS

作　　者：杨杰杰 徐倩[1] 杨玉玲[2] 李奕璇 王彬[3] 侯琳[1] 李宁[1] YANG Jiejie;XU Qian;YANG Yuling;LI Yixuan;WANG Bin;HOU Lin;LI Ning(Department of Biochemistry and Molecular Biology, School of Basic Medicine, Qingdao University, Qingdao 266071, China)

机构地区：[1]青岛大学基础医学院生物化学与分子生物学系,山东青岛266071 [2]青岛大学附属医院感染科 [3]青岛大学电子信息学院

出　　处：《精准医学杂志》2020年第4期318-322,共5页Journal of Precision Medicine

基　　金：国家自然科学基金(81600470);中国博士后科学基金特别资助项目(2016T90612);中国博士后科学基金面上资助项目(2015M57074);青岛市应用基础研究计划青年专项(19-6-2-59-cg)。

摘　　要：目的探讨内质网跨膜蛋白EVA1A在肝细胞癌(HCC)中的表达及对人肝细胞癌Huh7细胞生物学行为的影响。方法采用实时荧光定量PCR和免疫组化法检测EVA1A在HCC患者癌组织和癌旁正常组织中的表达;通过Western blot检测肝细胞L02与HCC细胞系Huh7中EVA1A的表达量;HCC细胞Huh7过表达EVA1A后,通过MTT法、细胞划痕法、DAPI染色观察EVA1A对Huh7细胞增殖、迁移和凋亡的影响。结果EVA1A在HCC患者癌组织和HCC细胞系Huh7中的表达较癌旁正常组织和正常肝细胞L02均显著降低(t=33.23、17.98,P<0.01)。与对照组相比,实验组过表达EVA1A后细胞增殖水平显著减弱(F=39.74、44.50,P<0.01);细胞迁移能力明显下降(t=7.09,P<0.05);实验组细胞发生典型的凋亡细胞形态学改变,相对凋亡率明显增加(t=-16.43,P<0.05)。结论EVA1A在HCC患者和HCC细胞系Huh7中的表达均下调。EVA1A过表达可抑制Huh7细胞增殖和迁移,诱发细胞凋亡。Objective To investigate the expression of endoplasmic reticulum transmembrane protein Eva-1 homolog A(EVA1A)in hepatocellular carcinoma(HCC)and its effect on the biological behavior of HCC cell line Huh7.Methods Real-time fluorescence quantitative PCR and immunohistochemistry were used to measure the expression of EVA1A in the cancer tissues and adjacent normal tissues of HCC patients.The expression levels of EVA1A in normal liver cells(L02)and HCC cell line Huh7 were determined by Western blot.After EVA1A was overexpressed in Huh7 cells,we evaluated its effects on the proliferation,migration,and apoptosis of Huh7 cells by MTT assay,wound healing assay,and DAPI staining,respectively.Results The expression of EVA1A in the cancer tissues of HCC patients and in HCC cell line Huh7 was significantly lower than that in adjacent normal tissues and L02 cells(t=33.23,17.98,P<0.01).After overexpression of EVA1A,compared with the control group,the experimental group had significantly reduced cell proliferation(F=39.74,44.50,P<0.01)and cell migration ability(t=7.09,P<0.05),as well as typical morphological changes of apoptotic cells,with a significantly increased relative apoptosis rate(t=-16.43,P<0.05).Conclusion The expression of EVA1A in HCC patients and HCC cell line Huh7 is down-regulated.Overexpression of EVA1A can inhibit the proliferation and migration and induce the apoptosis of Huh7 cells.

关 键 词：癌 肝细胞 内质网 膜糖蛋白类 细胞系 肿瘤 细胞增殖 细胞运动 细胞凋亡 体外研究 

分 类 号：R735.7[医药卫生—肿瘤]