甲胎蛋白基因沉默对肝癌细胞迁移黏附能力的影响及其机制  

Effects of AFP gene silencing on migration and adhesion of hepatocellular carcinoma cells

作　　者：王莉 贺涛 冯世杰 万百顺 王效谦 张珉 张玲 韩风 WANG Li;HE Tao;FENG Shijie;WAN Baishun;WANG Xiaoqian;ZHANG Min;ZHANG Ling;HAN Feng(Henan Tumor Hospital Affiliated to Zhengzhou University,Zhengzhou 450008,China)

机构地区：[1]郑州大学附属肿瘤医院/河南省肿瘤医院,郑州450008

出　　处：《山东医药》2018年第47期11-14,共4页Shandong Medical Journal

基　　金：河南省基础与前沿研究项目(082300450110)

摘　　要：目的观察甲胎蛋白(AFP)基因沉默对肝癌细胞迁移黏附能力的影响,并探讨其作用机制。方法将肝癌细胞系EGHC-9901分为3组,实验组转染p Silencer3. 0-H1-AFP质粒以沉默AFP基因表达、载体对照组转染空载体、空白对照组未做任何处理,采用Transwell迁移实验检测各组细胞迁移能力,采用细胞聚集实验检测各组细胞同质黏附能力,采用层黏连蛋白黏附实验检测各组细胞异质黏附能力,采用Western blotting法检测各组细胞黏附分子E-钙黏蛋白、CD44V6、整合素β1、整合素α5、α-连环素、β-连环素、CD15。结果实验组、载体对照组、空白对照组每个镜下穿膜细胞数分别为(132±16)个、(335±21)个、(371±32)个,实验组与载体对照组、空白对照组相比,P均<0. 05。振摇时间40 min时,实验组、载体对照组、空白对照组细胞聚集指数(AI)分别为0. 530±0. 020、0. 223±0. 017、0. 240±0. 012,实验组与载体对照组、空白对照组相比,P均<0. 05;振摇时间60 min时,实验组、载体对照组、空白对照组AI分别为0. 632±0. 031、0. 322±0. 021、0. 376±0. 019,实验组与载体对照组、空白对照组相比,P均<0. 05。培养60 min时,实验组、载体对照组、空白对照组OD值分别为0. 157±0. 008、0. 347±0. 026、0. 363±0. 039,实验组与载体对照组、空白对照组相比,P均<0. 05;培养90 min时,实验组、载体对照组、空白对照组OD值分别为0. 139±0. 010、0. 476±0. 023、0. 503±0. 045,实验组与载体对照组、空白对照组相比,P均<0. 05。实验组、载体对照组、空白对照组整合素β1蛋白表达量分别为0. 337、0. 078、0. 051,实验组与载体对照组、空白对照组相比,P均<0. 05;实验组、载体对照组、空白对照组CD15蛋白表达量分别为0. 221、0. 629、0. 631,实验组与载体对照组、空白对照组相比,P均<0. 05。结论 AFP基因沉默可抑制肝癌细胞迁移,促进肝癌细胞的同质黏附能力,抑制肝癌细胞异质�Objective To observe the effects of AFP gene silencing on migration and adhesion of hepatocellular carcinoma cell line EGHC-9901 and its mechanism.Methods EGHC-9901 cells were divided into three groups:the experimental group which was transfected with AFP-siRNA,the vector control group which was transfected with empty vector,and the blank control group without treatment.Transwell assay was applied to detect cell migration ability;the cell aggregation assay was applied to evaluate alterations of homogeneous adhesion ability whereas Laminin adhesion assay was used to assess its heterogeneous adhesion ability;Western blotting was used to detect the expression of migration and adhesion-related proteins E-cadherin,CD44V6,CD15,integrinβ5,β-catenin,andα-catenin.Results Transwell assay demonstrated more cells of the experimental group(132±16)that migrated through the polycarbonate membrane than those of the vector control group(335±21)and blank control group(371±32)(P<0.05).Cell aggregation assay witnessed markedly higher aggregation index of the experimental group(0.530±0.020 and 0.632±0.031),versus that of the vector control group(0.223±0.017 and 0.322±0.021)and the blank cotrol group(0.240±0.012 and 0.376±0.019)when the incubation time was 40 min and 60 min,with significant difference(all P<0.05).When we cultured cells for 60 min,the OD values of the experimental group,vector control group and blank control group were 0.157±0.008,0.347±0.026,and 0.363±0.039,respectively,with statistically significant difference(all P<0.05).At 90 min,the OD values of the experimental group,the vector control group and the blank control group were 0.139±0.010,0.476±0.023,and 0.503±0.045,respectively,with statistically significant difference(all P<0.05).The expression levels of integrinβ1 protein in the experimental group,vector control group and blank control group were 0.337,0.078,and 0.051,respectively;the expression levels of CD15 protein in the experimental group,vector control group and blank control group were 0

关 键 词：甲胎蛋白 肝肿瘤 肝癌 整合素Β1 细胞黏附分子15 

分 类 号：R73-3[医药卫生—肿瘤]