摘要
目的:研究转化生长因子(TGF-β1)诱导的上皮间质转换(EMT)对人胆囊癌细胞系GBC-SD中侧群(SP)细胞表达的影响.方法:在体外培养的GBC-SD中加入TGF-β1后,相差倒置显微镜下观察细胞形态学变化,Westernblot检测E-cadherin、Vimentin的表达改变,采用流式细胞术检测TGF-β1处理GBC-SD前、后SP细胞亚群的表达比例;撤去TGF-β1后,检测细胞形态学变化、E-cadherin和Vimentin的表达改变以及SP细胞亚群的表达比例.结果:人胆囊癌细胞系GBC-SD中存在SP细胞,其比例约为0.87%;TGF-β1可诱导GBC-SD细胞向间质型细胞形态转化,并上调间质标记蛋白Vimentin和下调上皮标记蛋白E-cadherin的表达,TGF-β1处理后GBC-SD细胞中SP细胞亚群的表达比例增加至2.3%;撤去TGF-β1后,发生间质型转化的GBC-SD向上皮细胞形态转化,间质标记蛋白Vimentin表达下调和上皮标记蛋白E-cadherin表达上调,SP细胞逐渐恢复至TGF-β1诱导前的水平.结论:TGF-β1诱导的可逆性EMT上调人胆囊癌细胞系GBC-SD中SP细胞亚群的表达,促进了肿瘤的衍进.
AIM: To investigate the impact of transforming growth factor-β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) on side population cells in human gallbladder cancer cell line GBC-SD. METHODS: GBC-SD cells were allowed to adhere and then incubated with human recombinant TGF-β1 to induce EMT in vitro. Morpho-logical alterations were examined by phasecontrast microscopy, and the expression of EMT-related proteins was detected by Western blot. The percentage of SP cells in GBC-SD cells with and without TGF-β1 was analyzed by flow cytometry. After the removal of TGF-β1, cellular morphological alterations, the expression of EMT-related proteins, and the percentage of SP cells were detected again. RESULTS: SP cells accounted for approximately 0.87% of GBC-SD cells. TGF-β1 induced morphological transition from epithelial to mesenchymal cells. After treatment with TGF-β1, the expression of epithelial cell marker E-cadherin was decreased, that of mesenchymal cell marker vimentin was increased, and the percentage of SP cells was increased to 2.3%. After removal of TGF-β1, transitioned GBC-SD cells regained the epithelial phenotypes and restored the SP abundance. CONCLUSION: TGF-β1-induced reversible EMT increases the percentage of SD cells in human gallbladder cancer cell line GBC-SD, suggesting that EMT may promote tumor progression by augmenting the abundance of SP cells.
出处
《世界华人消化杂志》
CAS
北大核心
2011年第16期1680-1685,共6页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30772127~~
关键词
胆囊癌
侧群细胞
转化生长因子β1
上皮间质转化
Human gallbladder cancer
Side population cells
Transforming growth factor-β1
Epithe- lial-mesenchymal transition