转化生长因子β1对间皮细胞上皮-间质表型转化的调控及对胃癌腹膜转移的影响  

Role of transforming growth factor-β1 in epithelial-mesenchymal transition of mesothelial cells and its effect on peritoneal metastasis of gastric cancer

作　　者：吕志栋[1] 孔滨[1] 刘相萍[2] 李福年[1] 王海波[1] 徐惠绵[3] 

机构地区：[1]青岛大学医学院附属医院乳腺外科,266003 [2]青岛大学医学院附属医院中心实验室,266003 [3]中国医科大学附属第一医院肿瘤外科

出　　处：《中华胃肠外科杂志》2015年第1期54-57,共4页Chinese Journal of Gastrointestinal Surgery

基　　金：国家自然科学基金（81302290）

摘　　要：目的：观察转化生长因子β1（TGF-β1）对腹膜间皮细胞上皮-间质表型转化的调控及对胃癌细胞腹膜转移的影响。方法 TGF-β1作用人腹膜间皮细胞系HMrSV5，倒置显微镜下观察间皮细胞形态学变化。采用Western-blot检测上皮细胞表型蛋白（细胞角蛋白和E-钙黏素）、间皮细胞表型蛋白（α-SMA和弹性蛋白）及Smad2蛋白水平的表达情况。采用黏附试验观察表型转化间皮细胞对胃癌细胞系HSC-39黏附的影响。间皮细胞（8×104／孔）与胃癌细胞系HSC-39（4×104／孔）共培养，采用体外Transwell侵袭实验观察腹膜微环境变化对胃癌细胞侵袭能力的影响。结果 TGF-β1作用腹膜间皮细胞24 h后部分细胞转变为狭长型，72 h后间皮细胞转变为典型纤维细胞样外观。TGF-β1作用间皮细胞可诱导弹性蛋白和α-SMA表达上调，细胞角蛋白和 E-钙黏素表达下降，并且呈现时间依赖性变化（P＜0．05）。 TGF-β1作用15 min后，间皮细胞内磷酸化Smad2表达开始升高，30 min达到顶峰，较对照组增加432％（P＜0．01）；但总Smad2表达则无明显变化（P＞0．05）。 TGF-β1作用72 h后，间皮细胞与HSC-39胃癌细胞的黏附率较对照组增加（146±17）％（P＜0．05）。胃癌细胞与TGF-β1刺激的间皮细胞共培养48 h后，平均每视野转移癌细胞数目为61．1±11．4，较对照组（31．9±8．1）明显增多（P＜0．05）。结论 TGF-β1能够诱导间皮细胞向成纤维细胞样转化，Smad2信号转导通路在间皮细胞表型转化中发挥重要作用；而这种腹膜微环境变化可增强胃癌细胞的黏附和侵袭能力，为癌细胞转移播散提供适宜的“土壤”环境。Objective To elucidate the role of transforming growth factor-beta1 （TGF-β1） in epithelial-mesenchymal transition of mesothelial cells and peritoneal metastasis of gastric cancer. Methods HMrSV5 cells, a human peritoneal mesothelial cell line, were incubated with TGF-β1, and their morphological changes were observed by phase contrast microscopy. Expressions of α-smooth muscle actin （α-SMA）, vimentin, cytokeratin, E-cadherin, phosphorylated-Smad2 and Smad2 were examined by Western blotting. After fibroblastic-like mesothelial cells were co-incubate with HSC-39 cells （gastric cancer cell line）, the adhesion and invasion potential of HSC-39 were evaluated by adhesion and invasion assay in vitro. Results Few mesothelial cells converted to spindle fibroblast-like morphology for 24 h, and remarkable phenotypic changes were observed at 72 h of TGF-β1 activation. TGF-β1 could induce α-SMA and vimentin expression, and down-regulate cytokeratin and E-cadherin expression in mesothelial cells （P0.05）. The percentage of HSC-39 gastric cancer cells adhered were significantly increased as compared to the control. When the mesothelial cells were treated by TGF-β1 for 72 h, the increased adhesion percentage was （146 ±17）%（P〈0.05）. After fibroblastic-like mesothelial cells co-incubated with HSC-39 cells for 48 h , more cancer cells [（61.1 ±11.4） cells/view field] invaded the coated membrane as compared to the control group [（31.9±8.1） cells/view field] （P〈0.05）. Conclusion TGF-β1 can induce the transition of mesothelial cells into myofibroblasts and Smad2 signal pathway may play a role in this transition , which is associated with increased adhesion and invasiveness of gastric cancer cells , and provides favorable environment for the dissemination of gastric cancer.

关 键 词：胃肿瘤 上皮-间质转化 间皮细胞 转化生长因子Β1 

分 类 号：R735.2[医药卫生—肿瘤]