蟾毒灵对TGF-β1诱导的HCT-116细胞上皮间质转化作用的影响  

The influence of Bufalin on TGF-β1-induced epithelial mesenchymal transition in HCT-116 cells

作　　者：赵汝楠 于卉[1] 石晓静[1] 邱艳艳[1] 钱雅新[1] 殷佩浩[1] 

机构地区：[1]上海中医药大学附属普陀医院普外科,上海200062

出　　处：《外科研究与新技术》2015年第4期217-222,共6页Surgical Research and New Technique

基　　金：上海中医药大学"杏林学者"(B-X-72);中西医结合一流学科创新基金项目(B-X-73);上海市普陀区卫计委"315"工程人才培养计划学科带头人后备人才(14Q-RC-08)

摘　　要：目的观察蟾毒灵(Bufalin)对大肠癌HCT-116细胞上皮间质转化作用的影响及其可能的机制。方法使用TGF-β1诱导大肠癌HCT-116细胞发生上皮间质转化(epithelial-mesenchymal transition,EMT),制备EMT模型。实验分为空白对照组、模型组、蟾毒灵组。72 h后,应用光学显微镜、侵袭实验、迁移实验观察蟾毒灵对HCT-116细胞形态学、侵袭与迁移能力的影响,Western blot检测EMT相关蛋白E-cadherin、Vimentin及β-catenin表达,免疫荧光法观察β-catenin蛋白分布。结果模型组较空白组细胞明显伸长变窄且细胞间连接相对疏松。与模型组相比,蟾毒灵组部分细胞呈不典型鹅卵石样上皮细胞改变,部分细胞拉长变窄,细胞间连接较紧密;72 h穿膜侵袭细胞数较模型组明显减少(110.00±26.46 vs.413.33±41.63,P<0.05),72 h迁移细胞数亦较模型组明显减少(507±38.16 vs.898±49.49,P<0.05),两者与空白对照组均无差异。Western blot检测显示:模型组较空白对照组上皮标志物E-cadhrin下降,间质标志物Vmentin上升;蟾毒灵组则较模型组E-cadhrin上调,Vimentin下调,β-catenin表达无差异。免疫荧光观察示,模型组β-catenin较空白对照组从细胞膜、胞质移至胞核内,蟾毒灵组β-catenin则较模型组集中于细胞质,胞核表达较少。结论 TGF-β1可诱导结肠癌HCT-116细胞EMT;蟾毒灵可能通过抑制β-catenin核移位、阻止HCT-116细胞EMT发挥抑制HCT-116细胞侵袭、迁移的作用。Objective To explore the influence of Bufalin on epithelial mesenchymal transition （EMT） in colorectal cancer HCT-116 cells and its possible mechanism. Methods EMT models of colorectal cancer HCT-116 cells were prepared by TGF-β1. Blank control group, model group and bufalin group were divided, and interventions were conducted. After 72 h of intervention, the morphological changes, invasion and migration capabilities, expression of EMT-related proteins including E-cadherin, Vimentin and β-catenin, and β-catenin protein distribution of HCT-116 cells in each group were determined by using light microscope, Transwell-invasion and migration tests, Western blotting and immunofluorescence method, respectively. Results Cells in model group became elongated and spindle, with relatively lax connections between ceils when compared with blank control group, and some cells in Bufalin group were elongated and narrow, with close cell connections when compared with model group. The number of cell permeation in Bufalin group （110.00±26.46） was significantly smaller than that in model group （413.33±41.63） （P〈0.05）, and the number of cell migration in Bufalin group was significantly decreased when compared with model group （507 ±38.16 vs. 898 ± 49.49, P〈0.05）, whereas there was no significant difference in the numbers of cell permeation and migration between blank control group and Bufalin group. Compared with model group, the expression of E-cadherin was increased, and the expression of Vimentin was decreased in Bufalin group. Compared with blank control group, the expression of E-cadherin was decreased, and the expression of Vimentin was increased in model group. However, there was no significant difference in β-catenin expression among groups. The β-catenin expression moved from cell membrane and cytoplasm to nucleus in model group when compared with blank control group, and there was more β-catenin expression in cytoplasm than nucleus in Bufalin group when compared with model group. Conclusion

关 键 词：大肠癌 蟾毒灵 上皮间质转化 转化生长因子-Β1 侵袭 迁移 

分 类 号：R735.34[医药卫生—肿瘤]