摘要
目的探讨微小RNA-150-5p(miR-150-5p)是否可靶向锌指E盒结合蛋白1(ZEB1)调控子宫内膜癌(EC)细胞上皮间质转化(EMT)进而影响癌细胞的恶性生物学行为。方法RT-qPCR技术检测正常子宫内膜和EC组织中、子宫内膜上皮细胞系hEEC及EC细胞系Ishikawa和HEC-1-A中miR-150-5p相对表达量。过表达miR-150-5p,MTT法、菌落形成实验、伤口愈合实验和Transwell实验分别评估Ishikawa细胞活力、克隆形成、迁移及侵袭能力;双荧光素酶报告基因实验验证miR-150-5p与ZEB1的靶向关系;RT-qPCR检测miR-150-5p及ZEB1 mRNA相对表达量;Western blot技术检测ZEB1、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)及基质金属蛋白酶9(MMP-9)蛋白表达量。结果与正常子宫内膜组织比较,EC组织中miR-150-5p相对表达量降低(P<0.05);与hEEC细胞比较,HEC-1-A细胞和Ishikawa细胞中miR-150-5p相对表达量降低(P<0.05),Ishikawa细胞中最低(P<0.05)。与空白组比较,miR-150-5p mimics组细胞490 nm处吸光度值、细胞菌落数、迁移数和侵袭数、ZEB1 mRNA和蛋白相对表达量及N-cadherin、Vimentin和MMP-9蛋白相对表达量显著降低,miR-150-5p相对表达量、E-cadherin蛋白相对表达量显著升高(P<0.05)。经生物信息学分析,ZEB1被预测为miR-150-5p的潜在靶基因。结论miR-150-5p可靶向ZEB1抑制癌细胞的恶性生物学行为,其作用机制可能与调控EC细胞EMT进展有关。
Objective To investigate whether microRNA-150-5p(miR-150-5p)can target zinc finger E-box binding protein 1(ZEB1)to regulate epithelial mesenchymal transformation(EMT)of endometrial carcinoma(EC)cells,thereby affecting the malignant biological behavior of cancer cells.Methods The relative expression of miR-150-5p in normal endometrium and EC tissues,endometrial epithelial cell lines hEEC,EC cell lines Ishikawa and HEC-1-A were detected by RT-qPCR.Overexpression of miR-150-5p,MTT assay,colony formation test,wound healing test and Transwell test were used to evaluate the viability,clonal formation,migration and invasion of Ishikawa cells.Double luciferase reporter gene experiment verified the targeting relationship between miR-150-5p and ZEB1.The relative expression of miR-150-5p and ZEB1 mRNA was detected by RT-qPCR.The expressions of ZEB1,E-cadherin,N-cadherin,vimentin and matrix metalloproteinase-9(MMP-9)were detected by Western blot.Results Compared with normal endometrial tissue,the relative expression of miR-150-5p in EC tissue was decreased(P<0.05).Compared with hEEC cells,the relative expression of miR-150-5p in HEC-1-A cells and Ishikawa cells decreased(P<0.05),and that in Ishikawa cells was the lowest(P<0.05).Compared with the blank group,the absorbance value at 490 nm,the number of colony,migration and invasion,the relative expression of ZEB1 mRNA and protein,and the relative expression of N-cadherin,Vimentin and MMP-9 protein in miR-150-5p mimics group significantly decreased,while the relative expression of miR-150-5p and E-cadherin protein significantly increased(P<0.05).According to bioinformatics analysis,ZEB1 was predicted as a potential target gene of miR-150-5p.Conclusion miR-150-5p can target ZEB1 to inhibit the malignant biological behavior of cancer cells,and its mechanism may be related to the regulation of EMT progression of EC cells.
作者
张桂萍
韩立
彭丽
ZHANG Guiping;HAN Li;PENG Li(Nanyang First People's Hospital,Nanyang,473000)
出处
《实用癌症杂志》
2024年第6期878-882,890,共6页
The Practical Journal of Cancer
作者简介
通讯作者:韩立。
