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

Abstract

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. Key words: Gallbladder neoplasms; Cell line; Neoplasm metastasis