胆囊癌高侵袭细胞株构建和生物学特性的研究  

Establishment and biological characterization of gallbladder carcinoma cell sublines with higher metastatic potential ability

作　　者：檀占海 李茂岚[1] 谈竹君[1] 董平[1] 刘颖斌[1] 张学利 

机构地区：[1]上海交通大学医学院附属新华医院普外科普外科实验室上海交通大学医学院胆道疾病研究所,200092 [2]上海市奉贤区中心医院普外科

出　　处：《中华实验外科杂志》2014年第11期2371-2373,共3页Chinese Journal of Experimental Surgery

基　　金：国家高技术研究发展（863）计划资助项目（2012AA022606）;国家自然科学基金面上项目（81172029、81172026）

摘　　要：目的 建立具有高侵袭转移能力胆囊癌细胞株亚系（GBC-SD-I-2）并观察其生物特性.方法 利用Transwell侵袭小室从人胆囊癌细胞株（GBC-SD）筛选高转移侵袭能力GBC-SD-I-2,光镜下比较细胞形态;克隆形成试验及噻唑蓝法（MTT）检测细胞增殖能力变化;实时定量荧光聚合酶链反应（Real-time PCR）及Western blot检测与上皮-间充质转化（EMT）相关N-钙黏蛋白、β-连环蛋白（β-catenin）的表达变化.结果 成功筛选出高侵袭转移能力细胞亚系.克隆形成试验证明其增殖能力明显增强（克隆形成率：0.711 ±0.021比0.445 ±0.017,P＜0.01）、MTT法显示体外生长能力显著强于母系（P＜0.05）.Real-time PCR检测显示亚系细胞N-钙黏蛋白表达水平明显升高（0.881±0.018比0.102±0.02,P＜0.01）;β-catenin表达明显降低（0.625±0.041比0.275±0.052,P＜0.05）.Western blot检测亚系N-钙黏蛋白表达水平明显升高（0.758±0.012比0.156±0.011,P＜0.01）;β-catenin表达明显降低（0.243±0.014比0.72 ±0.011,P＜0.01）.结论 筛选的胆囊癌细胞亚群GBC-SD-I-2的侵袭转移能力明显增强.Objective To establish high transfer potential,high attack ability gallbladder carcinoma cell line （GBC-SD） subline and study its biological characteristics.Methods using Transwell chamber from gallbladder carcinoma cell line （GBC-SD） screening high transfer invasion ability gallbladder carcinoma subline GBC-SD-I-2,then compare cells form; Clone formation test and four methyl azo azole blue （MTT） method to detect the change of cell proliferation ability; Real-time polymerase chain reaction （Real-time PCR） and Western blotting method detection and epithelial mesenchymal transition （EMT） related N-cadherin,β-catenin protein expression change.Results The use of small room Transwell invasion from gallbladder carcinoma cell line were high transfer potential cell subsets and maternal form there are significant differences.Clone formation assay （0.711 ± 0.021 vs.0.445 ± 0.017,P 〈 0.01）,MTT method shows subline proliferation ability noticeably （P 〈 0.05）.Real-time PCR test showed that the group of cells N-cadherin expression level increased significantly （0.881 ± 0.018 vs.0.102 ± 0.020,P 〈 0.01）,β-catenin protein expression decreased obviously （0.625 ± 0.041 vs.0.275 ±0.052,P 〈 0.05）.Western blotting method showed N-cadherin expression level increased significantly （0.758 ± 0.012 vs.0.156 ± 0.011,P 〈 0.01） ; β-catenin protein expression decreased obviously （0.243 ± 0.014,0.720 ± 0.011,P 〈 0.01）.Conclusion Screening gallbladder carcinoma cell subsets GBC-SD-I-2 attacks,transfer ability enhanced obviously,to explore the gallbladder carcinoma transfer mechanism provides the ideal experimental tools.

关 键 词：胆囊肿瘤 细胞株 肿瘤转移 

分 类 号：R735.8[医药卫生—肿瘤]