跨膜emp24结构域蛋白4在肝癌患者肝组织中的表达及其对肝癌细胞生物学行为的影响  

Expression of the transmembrane emp24 domain-containing protein 4 in liver tissues of patients with hepatocellular carcinoma and its effects on biological behavior of hepatocellular carcinoma cells

作　　者：汪礼洋 黄伟[1] 吴书桢 马韬 刘肇修[1] 陆翠华[1] Wang Liyang;Huang Wei;Wu Shuzhen;Ma Tao;Liu Zhaoxiu;Lu Cuihua(Department of Gastroenterology,Affiliated Hospital of Nantong University,Nantong 226001,China;Clinical Medical Research Center,Affiliated Hospital of Nantong University,Nantong 226001,China)

机构地区：[1]南通大学附属医院消化内科,南通226001 [2]南通大学附属医院临床医学研究中心,南通226001

出　　处：《中华消化杂志》2022年第10期667-674,共8页Chinese Journal of Digestion

基　　金：国家自然科学基金面上项目(82070624);国家自然科学基金青年基金(82000497);江苏省卫生健康委科研重点项目(ZDB2020006);江苏省自然科学基金青年基金(BK20200965)。

摘　　要：目的检测跨膜emp24结构域蛋白4(TMED4)在肝癌患者肝组织中的表达情况,并初步探究TMED4基因对肝癌细胞增殖和迁移能力的影响及其相关分子机制。方法采用蛋白质印迹法和免疫组织化学染色检测肝癌患者癌组织、癌旁组织中TMED4的蛋白质表达水平,并分析其表达与患者的临床病理参数之间的相关性;分别通过细胞增殖实验、Transwell实验、划痕愈合实验和裸鼠皮下成瘤实验探究过表达或敲减TMED4基因对肝癌细胞体内外增殖、迁移和愈合能力的影响;通过通路分析初步探究TMED4基因调控肝癌细胞生物学行为的可能分子机制。统计学方法采用独立样本t检验、Mann-Whitney U检验和卡方检验。结果蛋白质印迹法结果显示,TMED4在肝癌组织中的蛋白质表达水平低于其对应的癌旁组织(0.52±0.29比0.83±0.22),差异有统计学意义(t=2.54,P=0.022)。免疫组织化学染色结果显示,TMED4在肝癌组织中的蛋白质表达水平低于其对应的癌旁组织(5.46±3.37比7.58±3.08),差异有统计学意义(t=3.49,P<0.001)。TMED4的蛋白质表达水平与患者是否发生肿瘤血管侵犯和巴塞罗那临床肝癌(BCLC)分期显著相关(χ^(2)=6.83、4.20,P=0.009、0.040)。细胞增殖实验结果显示,SMMC-7721细胞中TMED4过表达组细胞的光密度值低于对照组(1.38±0.05比2.37±0.08),HepG2细胞中TMED4敲减组细胞的光密度值高于对照组(0.76±0.04比0.54±0.01),差异均有统计学意义(t=18.23、8.85,均P<0.001)。Transwell实验结果显示,SMMC-7721细胞中TMED4过表达组的迁移细胞数少于对照组(286.30±13.01比439.70±12.34),HepG2细胞中TMED4敲减组的迁移细胞数多于对照组(249.00±6.00比160.00±6.56),差异均有统计学意义(t=14.81、17.34,均P<0.001)。划痕愈合实验结果显示,SMMC-7721细胞中TMED4过表达组的细胞愈合率低于对照组[(0.21±0.01)%比(0.45±0.01)%],HepG2细胞中TMED4敲减组的细胞愈合率高于对照组[(0.46±0.01)%比(0.Objective To examine the expression of transmembrane emp24 domain-containing protein 4(TMED4)in liver tissue of patients with hepatocellular carcinoma,and to investigate the effects of TMED4 gene on the proliferation and migration of hepatocellular carcinoma cells and related molecular mechanisms.Methods The expression of TMED4 at protein level in liver cancer tissue and paracancerous tissue of patients with hepatocellular carcinoma were detected by Western blotting and immunohistochemical stainning,and the correlation between its expression and clinicopathological features was analyzed.The effects of TMED4 overexpression or knockdown on proliferation,migration and healing ability of hepatocellular carcinoma cells in vitro and in vivo were determined by cell proliferation test,Transwell test,wound healing test and subcutaneous tumor formation in nude mice.The molecular mechanism of TMED4 in regulating the biological behavior of hepatocellular carcinoma cells was preliminarily explored by pathway analysis.Independent sample t test,Mann-Whitney U test and chi-square test were used for statistical analysis.Results The results of Western blotting showed that the expression of TMED4 at protein level in hepatocellular carcinoma tissue was lower than that in paracancerous tissue(0.52±0.29 vs.0.83±0.22),and the difference was statistically significant(t=2.54,P=0.022).The results of immunohistochemical examination indicated that the expression of TMED4 at protein level in liver cancer tissue was lower than that in paracancerous tissue(5.46±3.37 vs.7.58±3.08),and the difference was statistically significant(t=3.49,P<0.001).The expression of TMED4 at protein level was significantly correlated with vascular invasion and Barcelona clinic liver cancer stage(χ^(2)=6.83 and 4.20,P=0.009 and 0.040).The results of cell proliferation assay showed that the absorbance value of SMMC-7721 cells in TMED4 overexpression group was lower than that in control group(1.38±0.05 vs.2.37±0.08),while the optical density value of HepG2 in T

关 键 词：癌 肝细胞 TMED4 细胞增殖 细胞迁移 

分 类 号：R735.7[医药卫生—肿瘤]