KAI1通过调节TGF-β1/Smad2信号通路减弱上皮-间充质转化抑制胃癌细胞的侵袭和迁移  

KAI1 Suppresses invasion and metastasis of gastric cancer by regulating the epithelial-mesenchymal transition through the TGF-β1/Smad2 signaling pathway

作　　者：张嫄怡 李春鸣 张霞[1] 梁娜 Zhang Yuanyi;Li Chunming;Zhang Xia;Liang Na(Department of Pathology and Pathophysiology,Zhaoqing Medical College,Zhaoqing Guangdong,526020,China;Department of Pathology,the Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou,563000,China;Department of Embryo Education and Research,Zunyi Medical University,Zunyi Guizhou,563099,China)

机构地区：[1]肇庆医学高等专科学校、病理学与病理生理学教研室,广东肇庆526020 [2]遵义医科大学附属医院病理科,贵州遵义563000 [3]遵义医科大学、组织胚胎学教研室,贵州遵义563099

出　　处：《遵义医科大学学报》2023年第4期388-395,共8页Journal of Zunyi Medical University

基　　金：广东省教育厅青年创新人才自然科学资助项目(NO:2018GkQNCX108);肇庆市科技创新指导类项目(NO:201904031402)。

摘　　要：目的探讨KAI1通过调控TGF-β1/Smad2信号通路减弱上皮-间充质转化(EMT)对胃癌细胞侵袭、迁移的影响。方法将人胃癌SGC-7901细胞分为KAI1组、NC组、Control组,KAI1组转染携带KAI1基因的重组慢病毒、NC组转染空病毒、Control组不转染,采用Western blot和qPCR法检测3组细胞KAI1表达情况。用含TGF-β1的培养基培养3组细胞,分别为KAI1+TGF-β1,NC+TGF-β1和TGF-β1组,通过光镜观察各组细胞形态改变;Transwell细胞侵袭和划痕实验分别检测各组细胞的侵袭和迁移能力;Western blot法检测白细胞抑制因子2(Smad2)、磷酸化Smad2(p-Smad2)、E-钙黏蛋白(E-cadherin)和波形蛋白(Vimentin)表达水平。结果Western blot和qPCR提示KAI1成功转染人胃癌SGC-7901细胞。在镜下观察发现NC+TGF-β1组和TGF-β1组细胞形态改变明显,而KAI1+TGF-β1组细胞没有出现明显的伪足等结构。KAI1+TGF-β1组细胞侵袭数明显低于NC+TGF-β1组和TGF-β1组(P<0.05)。KAI1+TGF-β1组划痕区域相对距离变化小于NC+TGF-β1组和TGF-β1组;细胞迁移率明显低于NC+TGF-β1组和TGF-β1组(P<0.05)。Western blot提示KAI1+TGF-β1组E-cadherin蛋白表达水平明显高于NC+TGF-β1组和TGF-β1组(P<0.05),而Vimentin,Smad2,p-Smad2蛋白表达水平明显低于NC+TGF-β1组和TGF-β1组(P<0.05)。结论KAI1基因可以通过抑制TGF-β1/Smad2信号通路抑制EMT,从而抑制胃癌细胞的侵袭、迁移。Objective To investigate the effects of KAI1 on the invasion and metastasis of gastric cancer cells by regulating TGF-β1/Smad2 signaling pathway to attenuate epithelial-mesenchymal transition(EMT).Methods Human gastric cancer SGC-7901 cells were divided into KAI1 group,NC group and Control group.KAI1 group was transfected with recombinant lentivirus carrying KAI1 gene,NC group was transfected with null virus,and Control group was not transfected.Western blot and qPCR were used to detect KAI1 expression in three groups of cells.Three groups of human gastric cancer cells were treated with TGF-β1,and then designated as the KAI1+TGF-β1,NC+TGF-β1 and TGF-β1 groups.The morphological changes of the three groups were observed under light microscope;transwell migration and wound healing assays were then performed comparing the three groups of cells.The expression of Smad2,p-Smad2,E-cadherin and Vimentin were detected by western blot.Results Western blot and qPCR suggested that KAI1 was successfully transfected into human gastric cancer SGC-7901 cells.Under the microscope,it was found that the change of cell morphology in NC+TGF-β1 group and TGF-β1 group was significant,while the cell morphology of KAI1+TGF-β1 group didn’t show obvious pseudopodia and other structures.Transwell migration assays showed the number of invasion cells in KAI1+TGF-β1 group was obviously decreased(P<0.05).The relative distance change of scratch area in group KAI1+TGF-β1 was less than that in group NC+TGF-β1 and TGF-β1 group,and the percentage of wound closure was significantly decreased in KAI1+TGF-β1 group compared with NC+TGF-β1 and TGF-β1 group.Compared with NC+TGF-β1 and TGF-β1 group,levels of E-cadherin increased in the KAI1+TGF-β1 group,while levels of Vimentin,Smad2 and p-Smad2 were deceased in KAI1+TGF-β1 group(P<0.05).Conclusion Overexpression KAI1 could regulate gastric cancer migration,invasion by attenuating EMT via inhibition of TGF-β1/Smad2 signaling pathway.

关 键 词：胃癌 KAI1基因 TGF-β1/Smad2信号通路 上皮-间充质转化 侵袭 转移 

分 类 号：R361.3[医药卫生—病理学]