miR-7对Hp诱导胃上皮细胞自噬及EMT的影响  

Effects of miR-7 on Hp-induced autophagy and EMT in gastric epithelial cells

作　　者：曾茹 毛山山[2] ZENG Ru;MAO Shanshan(Department of Gstroenterology,Haikou People’s Hospital,Haikou 570000,Hainan,China;Department of Cancer Chemotherapy,Haikou People’s Hospital,Haikou 570000,Hainan,China)

机构地区：[1]海口市人民医院消化内科,海南海口570000 [2]海口市人民医院肿瘤化疗科,海南海口570000

出　　处：《中南医学科学杂志》2024年第2期177-181,共5页Medical Science Journal of Central South China

基　　金：海南省自然科学基金高层次人才项目(822RC862)。

摘　　要：目的探讨微小RNA-7(miR-7)对幽门螺杆菌(Hp)诱导的胃上皮细胞(GES-1)自噬及上皮细胞间质转化(EMT)的影响。方法体外将Hp悉尼株(SS1)与GES-1细胞共培养建立GES-1细胞转化模型;实验分为空白对照组(NC组)、Hp组、miR-7模拟物(mimics)组、Hp+miR-7 mimics组。qRT-PCR验证转染效果;MTT法、划痕实验、Transwell实验检测各组GES-1细胞增殖能力、迁移能力和侵袭能力;透射电镜下观察细胞自噬体;免疫印迹法检测各组GES-1细胞自噬相关蛋白[RM-9106微管相关蛋白1轻链3-Ⅰ(LC3-Ⅰ)、LC3-Ⅱ]及EMT相关蛋白[E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)]表达情况。结果与NC组比较,Hp组GES-1转化细胞增殖抑制率、p62蛋白、E-Cadherin蛋白水平降低(P<0.05),迁移率、侵袭数、自噬体数量及自噬泡/胞质总面积、LC3-Ⅱ/LC3-Ⅰ比值、N-Cadherin和Vimentin蛋白水平均升高(P<0.05),miR-7 mimics组GES-1细胞增殖抑制率、p62蛋白、E-Cadherin蛋白水平升高(P<0.05),迁移率、侵袭数、自噬体数量及自噬泡/胞质总面积、LC3-Ⅱ/LC3-Ⅰ比值、N-Cadherin和Vimentin蛋白水平均降低(P<0.05);与miR-7 mimics组比较,Hp+miR-7 mimics组转化细胞增殖抑制率、p62蛋白、E-Cadherin蛋白水平降低(P<0.05),迁移率、侵袭数、自噬体数量及自噬泡/胞质总面积、自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值、N-Cadherin和Vimentin蛋白水平均升高(P<0.05)。结论miR-7下调Hp诱导的GES-1细胞自噬,降低细胞侵袭及迁移能力,抑制EMT。Aim To investigate the effect of microRNA-7(miR-7)on Helicobacter pylori(Hp)-induced autophagy and epithelial mesenchymal transition(EMT)in gastric epithelial cells(GES-1).Methods The Hp Sydney strain(SS1)was co cultured with GES-1 cells in vitro to establish a GES-1 cell transformation model.The experiment was divided into blank control group(NC group),Hp group,miR-7 mimics group,and Hp+miR-7 mimics group.The transfection effect was verified by qRT-PCR.MTT assay,scratch assay and Transwell assay were used to detect the proliferation,migration and invasion of GES-1 cells in each group.Western blotting was used to detect the expression of autophagy related proteins(RM-9106 microtubule related protein 1 light chain 3-Ⅰ(LC3-Ⅰ),LC3-Ⅱ)and epithelial mesenchymal transformation(EMT)related proteins(E-cadherin,N-cadherin,Vimentin)in GES-1 cells in each group.Results Compared with the NC group,the Hp group showed a decrease in the proliferation inhibition rate,p62 protein,and E-Cadherin protein levels of GES-1 transformed cells(P<0.05),while the migration rate,invasion number,number of autophagosomes,total area of autophagic vesicles/cytoplasm,ratio of LC3-Ⅱ/LC3-Ⅰ,N-Cadherin and Vimentin protein levels increased(P<0.05).The miR-7 mimics group also showed an increase in the proliferation inhibition rate,p62 protein,and E-Cadherin protein levels of GES-1 cells(P<0.05),and the migration rate The number of invasions,the number of autophagosomes,the total area of autophagic vesicles/cytoplasm,the ratio of LC3-Ⅱ/LC3-Ⅰ,and the levels of N-Cadherin and Vimentin proteins all decreased(P<0.05);Compared with the miR-7 mimics group,the Hp+miR-7 mimics group showed a decrease in the inhibition rate of transformed cell proliferation,p62 protein levels,and E-Cadherin protein levels(P<0.05),while the migration rate,invasion number,number of autophagosomes,total area of autophagic vesicles/cytoplasm,ratio of autophagy related protein LC3-Ⅱ/LC3-Ⅰ,and levels of N-Cadherin and Vimentin proteins all increased(P<0.05).Conclus

关 键 词：微小RNA-7 幽门螺杆菌 胃上皮细胞GES-1 自噬 

分 类 号：R446.5[医药卫生—诊断学]