白藜芦醇通过Hippo-YAP信号通路在TGF-β1诱导胃癌细胞EMT过程中的作用及其机制

Effect of Resveratrol on TGF-β1 Induced EMT in Gastric Cancer Cells through Hi-YAP Signaling Pathway and Its Mechanism

目的探讨白藜芦醇(RSVL)通过Hippo-YAP信号通路在TGF-β1诱导胃癌细胞上皮间充质转化(EMT)过程中的作用及其机制。方法选取胃癌细胞SGC-7901为研究对象,首先将其随机分为空白组、RSVL低剂量组(5μM)、RSVL中剂量组(10μM)、RSVL高剂量组(20μM),通过细胞增殖(MTT)实验确定RSVL浓度,然后对细胞进行转染,分为si-NC组、TGF-β1+10μM RSVL组、TGF-β1+si-YAP组、TGF-β1+pcDNA3.1-YAP组、TGF-β1+10μM RSVL+pcDNA3.1-YAP组。采用MTT、细胞侵袭(Transwell)、划痕实验,分别检测细胞的增殖、侵袭和迁移;采用蛋白质印迹法(WB)和荧光定量PCR检测E-钙粘连蛋白(E-cadherin)、神经型钙黏附蛋白(N-cadherin)、波形蛋白(Vimentin)、Snali 1、HIP-PO/Yes相关蛋白(YAP)和mRNA。其次,选取24只裸鼠,将其分为模型组、RSVL组(10μM)、RSVL+pcDNA3.1组、RSVL+pcDNA3.1-YAP组,每组各六只。计算小鼠肿瘤的重量和体积;采用免疫组化检测Ki67蛋白。结果RSVL对细胞增殖有抑制作用(P<0.05);与si-NC组相比,其余各组侵袭细胞数、迁移率较低(P<0.05);与si-NC组相比,其余各组E-cadherin蛋白及mRNA表达较高,N-cadherin、Vimentin、Snali 1蛋白及mRNA以及YAP蛋白表达较低(P<0.05);与Model组相比,其余各组小鼠肿瘤的重量和体积均较低(P<0.05);与Model组比较,其余各组Ki67染色强度均显著减弱。结论RSVL可以通过抑制YAP相关通路的激活,来发挥抑制肿瘤细胞SGC-7901上皮间充质转化、增殖、迁移、侵袭的作用。

Objective To investigate the role of resveratrol(RSVL)in TGF-β1-induced epithelial mesenchymal transition(EMT)of gastric cancer cells through the Hippo-YAP signaling pathway and its mechanism.Methods Gastric cancer cells SGC-7901 were selected as the research object,firstly,they were randomly divided into blank group,RSVL low(5μM),medium(10μM),and high dose group(20μM),and the concentration of RSVL was determined by the cell proliferation(MTT)assay,and then the cells were transfected into the si-NC group,the TGF-β1+10μM RSVL group,the TGF-β1+si-YAP group,TGF-β1+pcDNA3.1-YAP group,and TGF-β1+10μM RSVL+pcDNA3.1-YAP group.MTT,cell invasion(Transwell),and scratch assay were used to detect cell proliferation,invasion,and migration,respectively;protein blotting(WB)and fluorescence quantitative PCR were used to detect E-cadherin,neurotypical cadherin(N-cadherin),vimentin,Snali 1,HIP-PO/Yes-associated protein(YAP)proteins and mRNA.second,24 nude mice were selected and divided into the model group,RSVL group(10μM),RSVL+pcDNA3.1 group,and RSVL+pcDNA3.1-YAP group,with six mice in each group.The weight and volume of mouse tumors were calculated;Ki67 protein was detected by immunohistochemistry.Results RSVL had an inhibitory effect on cell proliferation,P<0.05;compared with the si-NC group,the number of invaded cells and migration rate were lower in the remaining groups,P<0.05;compared with the si-NC group,the expression of E-cadherin protein and mRNA was higher in the remaining groups,and the expression of N-cadherin,Vimentin,Snali 1 protein and mRNA,and YAP protein expression was lower,P<0.05;the weight and volume of mice were lower in the remaining groups compared with the Model group,P<0.05;and the intensity of Ki67 staining was significantly weaker in the remaining groups compared with the Model group.Conclusion RSVL can inhibit the epithelial mesenchymal transformation,proliferation,migration and invasion of SGC-7901 cells by inhibiting the activation of YAP-related pathway.