文献类型：期刊文献

中文题名：miR-335-5p调控TGF-β/Smad信号通路抑制胃癌细胞EMT及侵袭和迁移

英文题名：miR-335-5p regulates TGF-β/Smad signaling pathway to inhibit EMT,invasion and migration of gastric cancer cells

作者：宁月[1,2,3];陈凤琴[1];赵雪灵[1];邵利华[2,3];王蒙[2,3];王宏伟[1];李海龙[1,2,3]

第一作者：宁月

机构：[1]甘肃中医药大学附属医院,甘肃兰州730000;[2]甘肃中医药大学第一临床医学院内科学教研室,甘肃兰州730000;[3]甘肃中医方药挖掘与创新转化重点实验室,甘肃兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2024

卷号：32

期号：22

起止页码：4228

中文期刊名：现代肿瘤医学

外文期刊名：Journal of Modern Oncology

基金：甘肃省卫生行业计划项目(编号:GSWSKY2022-25);甘肃中医药大学附属医院院内创新基金项目(编号:gzfy-2022-12)。

语种：中文

中文关键词：miR-335-5p;TGF-β/Smad通路;胃癌;EMT

外文关键词：miR-335-5p;TGF-β/Smad;gastric cancer;EMT

摘要：目的:基于TGF-β/Smad信号通路探讨上调微小RNA(miR)-335-5p对胃癌细胞侵袭、迁移及EMT分子表达的影响。方法:使用miR-335-5p慢病毒载体感染胃癌细胞,通过嘌呤霉素筛选构建稳转细胞株,采用划痕和Transwell实验观察上调miR-335-5p对胃癌细胞侵袭和迁移能力的影响。用RT-qPCR实验和Western-blot实验检测上调miR-335-5p对胃癌细胞TGF-β信号通路中关键基因和上皮间质转化(EMT)相关基因的mRNA和蛋白表达量的影响。结果:划痕结果显示,与阴性对照组相比,上调miR-335-5p后,两株胃癌细胞的愈合率明显降低(P<0.05,P<0.01)。Transwell迁移和侵袭实验结果显示,两株胃癌细胞miR-335-5p过表达组穿过小室的细胞数相对于阴性对照组明显降低(P<0.05,P<0.01)。RT-qPCR和Western-blot实验结果显示,两株胃癌细胞miR-335-5p过表达组的TGF-β1、TGF-β2、TGF-βR2、p-Smad2、p-Smad3和Smad4蛋白表达量明显低于阴性对照组(P<0.05,P<0.01)。miR-335-5p过表达后,两株胃癌细胞的α-平滑肌肌动蛋白(α-SmA)、I型胶原(Collagen 1)、N-钙黏着蛋白(N-cadherin)、细胞角蛋白(Cytokeratin18)、Claudin7蛋白(Claudin7)、闭锁小带蛋白-1(ZO-1)、波形蛋白(Vimentin)、锌指转录因子1(Snail1)、锌指转录因子2(Snail2)、纤连蛋白1(FN1)、Twist相关蛋白2基因(Twist2)的mRNA和蛋白表达量明显低于阴性对照组(P<0.05,P<0.01),E-钙黏着蛋白(E-cadherin)的mRNA和蛋白表达量高于阴性对照组(P<0.05,P<0.01)。结论:过表达miR-335-5p通过TGF-β/Smad信号通路调控胃癌细胞的EMT,从而抑制胃癌细胞的侵袭和迁移。
Objective:To investigate the effects of upregulation of microRNA(miR)-335-5p on the invasion,migration and the expressions of EMT markers in gastric cancer cells via TGF-β/Smad signaling pathway.Methods:Gastric cancer cells were infected with miR-335-5p lentiviral vector,and screened by puromycin to establish a stable cell line.The effects of up-regulated miR-335-5p on the invasion and migration of gastric cancer cells were observed by scratch and Transwell experiments.The effects of up-regulation of miR-335-5p on mRNA and protein expression levels of key genes of TGF-βsignaling pathway and the genes related to epithelial interstitial transition(EMT)in gastric cancer cells were detected by RT-qPCR and Western-blot assay.Results:The scratch results showed that compared with the negative control group,the healing rate of the two gastric cancer cells was significantly decreased after up-regulation of miR-335-5p(P<0.05,P<0.01).The results of Transwell migration and invasion experiments showed that the number of cells passing through the chamber of the two gastric cancer cell lines in the miR-335-5p overexpression group was significantly lower than that in the negative control group(P<0.05,P<0.01).The experiments results showed that the expressions of TGF-β1,TGF-β2,TGF-βR2,p-Smad2 and p-Smad3 and Smad4 protein in miR-335-5p upregulation group were significantly lower than the negative control group(P<0.05,P<0.01).The expressions of a-SmA,Collagen1,N-cadherin,Cytokinetin18,Claudin7,ZO-1,Vimentin,Snail1,Snail2,FN1,Twist2 mRNA and protein in the miR-335-5p overexpression group was significantly lower than the negative control group(P<0.05,P<0.01),while E-cadherin mRNA and protein levels higher than the negative control group(P<0.05,P<0.01).Conclusion:Overexpression of miR-335-5p regulates EMT via TGF-β/Smad signaling pathway,thereby inhibiting the invasion and migration of gastric cancer cells.