高、低表达miR-296-5p的肿瘤细胞外泌体对食管鳞癌细胞迁移和侵袭的调控作用  

Regulation of tumor cell exosomes with high and low expression of miR-296-5p on migration and invasion of ESCC cells

作　　者：陈河山[1] 许荣誉[1] 陈劭赓[1] 傅景梁 陈泓波 杜祥昆 何荣琦[1] CHEN Heshan;XU Rongyu;CHEN Shaogeng;FU Jingliang;CHEN Hongbo;DU Xiangkun;HE Rongqi(Department of Thoracic Surgery,Quanzhou First Hospital,Fujian Medical University,Quanzhou 362000,China;不详)

机构地区：[1]福建医科大学附属泉州第一医院胸外科,福建泉州362000 [2]福建医科大学第三临床医学院

出　　处：《山东医药》2024年第5期44-48,共5页Shandong Medical Journal

基　　金：福建省卫健委医学创新课题项目(2019-CX-46)。

摘　　要：目的探讨高、低表达miR-296-5p的肿瘤细胞外泌体对食管鳞癌细胞(ESCC)迁移和侵袭的调控作用及其机制。方法提取转染miR-296-5p过表达和miR-296-5p干扰及其阴性对照物(NC)后的ESCC细胞外泌体,并采用透射电镜与Western blotting法进行鉴定;将ESCC细胞分为三组,干扰组、过表达组分别加入miR-296-5p干扰或过表达细胞的外泌体培养,NC组加入NC转染细胞的外泌体培养48 h;取各组细胞,MTT法检测细胞活力、Transwell法检测细胞侵袭迁移能力,RT-PCR、Western blotting法检测细胞黏附分子3(CADM3)、CADM1、CADM4、钙黏蛋白E(E-cadherin)及血管内皮细胞生成浸润相关蛋白CD31、CD44。结果细胞活力、迁移和侵袭细胞数过表达组>NC组>干扰组(P均<0.05)。细胞中CADM1、CADM4、E-cadherin基因及蛋白表达水平干扰组>NC组>过表达组,CADM3、CD31、CD44基因及蛋白表达水平过表达组>NC组>干扰组(P均<0.05)。结论高表达miR-296-5p的肿瘤细胞外泌体可能通过调控CADM、E-cadherin、CD31、CD44等基因表达提高ESCC细胞的侵袭、迁移能力。Objective To explore the regulatory effect and mechanism of tumor cell exosomes with high and low expression or miR-296-5p on the metastasis of esophageal squamous cell carcinoma(ESCC)cells.Methods The exosomes of ESCC cells transfected with miR-296-5p overexpression,miR-296-5p interference and its negative control(NC)were extracted,and identified by transmission electron microscopy and Western blotting.ESCC cells were divided into three groups;cells in the interference group and the overexpression group were cultured with exosomes of miR-296-5p interference and overexpression cells,respectively;cells in the NC group were cultured with exosomes of NC-transfected cells for 48 h.Cells were collected from each group,cell viability was detected by MTT,and cell migration and invasion were detected by Transwell assay.The expression levels of recombinant cell adhesion molecule-3(CADM3),CADM1,CADM4,E-cadherin,platelet-endothelial cell adhesion molecule-1(CD31)and CD44 were detected by RT-PCR and Western blotting.Results The cell viability,the number of migration cells and the number of invading cells were as follows:the overexpression group>NC group>interference group,and there were significant differences(all P<0.05).The expression levels of CADM1,CADM4 and E-cadherin were as follows:the interference group>NC group>overexpression group,and there were significant differences between groups;the expression levels of CADM3,CD31 and CD44 were higher in the overexpression group than in the NC group,which were higher than those in the interference group,and the differences were statistically significant(all P<0.05).Conclusion Exosomes with high expression of miR-296-5p may promote the invasion and migration abilities of ESCC cells by regulating the expression levels of CADM,E-cadherin,CD31,CD44,and other genes.

关 键 词：微小核糖核酸296-5p 食管鳞癌 外泌体 细胞侵袭 细胞迁移 

分 类 号：R735.1[医药卫生—肿瘤]