TGF-β1诱导的上皮间质转化对肝癌细胞系中边缘群细胞的调控  

Regulation of side population cells by TGF-β1-induced epithelial-mesenchymal transition in hepatocellular carcinoma cell lines SMMC-7721 and HepG2

作　　者：王波[1] 夏羽佳[1] 晏维[1] 夏丽敏[1] 刘梅[1] 田德安[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院消化内科,湖北省武汉市430030

出　　处：《世界华人消化杂志》2010年第23期2416-2421,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30873038;No.30800968~~

摘　　要：目的:观察转化生长因子β1(TGF-β1)诱导的上皮间质转化(EMT)对SMMC-7721中SP细胞的影响.方法:常规培养人肝癌细胞系SMMC-7721和HepG2,利用流式细胞术分析肝癌细胞系中SP细胞的含量并分选出SP细胞,Western blot检测ABCG2蛋白的表达.TGF-β1诱导肝癌细胞SMMC-7721,动态观察细胞形态学变化,Western blot和免疫荧光检测EMT相关指标钙粘蛋白(E-cadherin)的表达,通过流式细胞仪分析TGF-β1诱导前后SP细胞含量的变化.结果:利用流式细胞术成功建立了肝癌SP细胞分选体系.检测发现,SMMC-7721中SP细胞含量为4.3%,Western blot显示该SP细胞ABCG2蛋白表达明显上调.而HepG2中未能检测出SP细胞.TGF-β1诱导72h后SMMC-7721细胞发生上皮间质转化,Western blot和免疫荧光检测发现E-cadherin蛋白的表达下调,同时SP细胞含量也明显降低.结论:不同肝癌细胞系中SP细胞含量的差别可能与细胞系的遗传背景和性质不同有关.EMT可能抑制了肿瘤干细胞的增殖,促进肿瘤细胞向侵袭和转移的方向发展.AIM：To evaluate the influence of transforming growth factor-β1（TGF-β1）-induced epithelialmesenchymal transition（EMT） on side population cells in SMMC-7721.METHODS：The percentages of SP cells in SMMC-7721 and HepG2 cells were analyzed by flow cytometry.TGF-β1 was added to the cultured SMMC7721 and HepG2 cells to induce EMT in vitro.Seventy-two hours later,cell morphological changes were observed,the pro-tein expression of E-cadherin was detected by Western blot and immunofluorescence,and the changes in the percentages of SP cells were analyzed by flow cytometry.RESULTS：An experimental system for the selection of SP cells from hepatocellular carcinoma cells was successfully established.There were 4.3% of SP cells in SMMC-7721 cells,and the expression of ABCG2（ATP-binding cassette superfamily G member 2） was up-regulated in these SP cells.No SP cells were detected in HepG2 cells.After exposure to TGF-β1 for 72 h,SMMC-7721 cells showed significant morphological changes and reduced E-cadherin expression and SP cell percentage.CONCLUSION：TGF-β1 is able to induce EMT and reduce the percentage of side population cells in hepatocellular carcinoma cell lines,suggesting that EMT may promote the invasion and metastasis of carcinoma cells by suppressing the proliferation of cancer stem cells.

关 键 词：边缘群细胞 转化生长因子Β1 上皮间质转化 肿瘤干细胞 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]