CAJ | 학술논문

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.
목적 건립구유고침습전이능력담낭암세포주아계(GBC-SD-I-2)병관찰기생물특성.방법 이용Transwell침습소실종인담낭암세포주(GBC-SD)사선고전이침습능력GBC-SD-I-2,광경하비교세포형태;극륭형성시험급새서람법(MTT)검측세포증식능력변화;실시정량형광취합매련반응(Real-time PCR)급Western blot검측여상피-간충질전화(EMT)상관N-개점단백、β-련배단백(β-catenin)적표체변화.결과 성공사선출고침습전이능력세포아계.극륭형성시험증명기증식능력명현증강(극륭형성솔:0.711 ±0.021비0.445 ±0.017,P＜0.01)、MTT법현시체외생장능력현저강우모계(P＜0.05).Real-time PCR검측현시아계세포N-개점단백표체수평명현승고(0.881±0.018비0.102±0.02,P＜0.01);β-catenin표체명현강저(0.625±0.041비0.275±0.052,P＜0.05).Western blot검측아계N-개점단백표체수평명현승고(0.758±0.012비0.156±0.011,P＜0.01);β-catenin표체명현강저(0.243±0.014비0.72 ±0.011,P＜0.01).결론 사선적담낭암세포아군GBC-SD-I-2적침습전이능력명현증강.
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.