B7-H4在肝癌组织中的表达及其对肝癌细胞侵袭、迁移的影响  

The expression of B7-H4 in liver cancer and its effect on cell invasion and migration

作　　者：何涛[1] 胡洪[1] 谢楠[2] 钱程 胡明[2] 张楚悦 柯恬恬 李春满[2] 付必莽[2] 苏莹珍 HE Tao;HU Hong;XIE Nan;QIAN Cheng;HU Ming;ZHANG Chuyue;KE Tiantian;LI Chunman;FU Bimang;SU Yingzhen(Department of Gastrointestinal Surgery,Yuxi People’s Hospital,Yuxi,Yunnan 653100,China;Department of Hepatobiliary and Pancreatic Surgery,Yunnan 650101,China;Department of Anesthesiology,The Second Affiliated Hospital of Kunming Medical University,Kunming,Yunnan 650101,China;Department of Laboratory Medicine and Preventive Medicine,Kunming University,Kunming,Yunnan 650214,China)

机构地区：[1]玉溪市人民医院胃肠外科,云南玉溪653100 [2]昆明医科大学第二附属医院肝胆胰外科,云南昆明650101 [3]昆明医科大学第二附属医院麻醉科,云南昆明650101 [4]昆明学院医学院公共卫生教研室,云南昆明650214

出　　处：《安徽医药》2019年第9期1774-1778,I0003,共6页Anhui Medical and Pharmaceutical Journal

基　　金：云南省教育厅科学研究基金项目（2019J0567）;云南省科学技术厅科技计划项目（2014FZ026）;云南省医疗卫生单位内设研究机构科研项目（2016NS248,2017NS281）;昆明学院引进人才科研项目（YJL18019）

摘　　要：目的探讨新型共刺激分子B7?H4在肝癌组织中的表达情况及其对肝癌细胞侵袭和迁移的影响。方法(1)收集2013年8月至2019年1月昆明医科大学第二附属医院肝癌组织32例;(2)筛选肝癌细胞系Hep3B,SMMC?7721,Huh?7,PLC/PRF/5等细胞的B7?H4表达情况;(3)筛选并培养高表达的该基因的肝癌细胞株,并使用预先制备好的慢病毒载体抑制该基因表达,将处于对数生长期的高表达的B7?H4肝癌细胞分为对照组、shRNA?1抑制组、shRNA?2抑制组,采用Western Blot(WB)法及实时荧光定量PCR法检测各组B7?H4的相对表达量;采用Tranwell细胞侵袭实验观察各组细胞侵袭能力;采用细胞划痕实验观察各组细胞迁移能力。结果在临床样本中,B7?H4在肝癌组织中的表达明显高于癌旁组织,且肝癌中B7?H4表达越高,病人越容易复发。HCCLM3的Western Blot相对表达量,对照组(5.32±1.03)、shRNA?1组(1.42±0.29)、shRNA?2组(2.06±0.68),shR?NA?1组、shRNA?2组均被抑制,shRNA?1组抑制明显(P<0.05)。HCCLM3对照组的细胞数为(365±21)、shRNA?1组(128±8);PLC/PRF/5组的细胞数为(267±44)、shRNA?1组(79±12)。与对照组相比较,shRNA?1组可明显抑制HCCLM3和PLC/PRF/5的细胞侵袭能力(P<0.01)。HCCLM3对照组的细胞迁移率为(82%±13%)、shRNA?1组(44%±6%);PLC/PRF/5对照组细胞迁移率为(77%±9%)、shRNA?1组(41%±6%)。与对照组比较,shRNA?1组明显抑制HCCLM3和PLC/PRF/5的细胞迁移能力(P<0.05)。转染抑制该基因后其相对表达量、细胞侵袭能力、迁移能力均下降。结论B7?H4有望成为新的肝癌预后标志物,下调肝癌细胞B7?H4的表达可以减弱肝癌细胞侵袭和迁移能力。Objective To investigate the relationship between the expression of novel co?stimulatory molecule B7?H4 in hepatocel?lular carcinoma and its pathology,and to explore its effect on the invasion and migration of hepatoma cells.Methods(1)Stained liver cancer sections were collected and analyzed(.2)The expression of B7?H4 in hepatoma cell lines Hep3B,SMMC?7721,Huh?7,PLC/PRF/5,HepG2,and HCCLM3 was screened(.3)Culture of a highly expressed B7?H4 hepatoma cell line and inhibition of the gene using a lentiviral vector.The highly expressed B7?H4 hepatoma cells in the logarithmic growth phase were divided into control group,shRNA?1 inhibition group and shRNA?2 inhibition group,and each group was detected by Western Blot(WB)and real?time fluorescence quantitative PCR.The relative expression level of H4;the invasion ability of each group was observed by Tranwell cell invasion assay;which were tested for the cell migration and invasion ability by scratch assay and Transwell migration assay.Re?sults The expression of B7?H4 in hepatocellular carcinoma was significantly higher than that of cancer in clinical samples.Periph?eral tissue,and the higher expression of B7?H4 in liver cancer,the more likely the patient relapsed.After transfection of the gene,its relative expression,cell invasion,and migration ability all decreased.Conclusion B7?H4 is expected to become a new prognos?tic marker of liver cancer.Down?regulating the expression of B7?H4 in hepatocellular carcinoma can weaken the invasion,migration,and tumorigenic potential of hepatoma cells.

关 键 词：肝癌 共刺激分子B7-H4 病理 细胞侵袭 细胞迁移 

分 类 号：R73[医药卫生—肿瘤]