摘要
目的分析miR-151-3p在子宫内膜异位症(EMS)患者子宫内膜组织中的表达水平及其对子宫内膜基质细胞(ESCs)迁移和侵袭的影响。方法收集87份EMS患者的在位子宫内膜组织(eEMS组)和异位子宫内膜组织(nEMS组),同时收集47份因子宫肌瘤等良性妇科疾病进行手术治疗的患者正常子宫内膜组织作为对照组,qRT-PCR法检测各组患者子宫内膜组织中miR-151-3p的表达水平。分离、鉴定、培养人ESCs细胞,再将miR-151-3p mimics、mimics NC、miR-151-3p inhibitors和inhibitors NC分别转染至ESCs细胞中,采用qRT-PCR检测各组细胞中miR-151-3p表达水平;CCK-8检测细胞增殖能力;Transwell检测细胞侵袭与迁移能力;Western blot检测各组细胞MMP-2和MMP-9以及上皮-间充质转化(EMT)标记物E-cadherin、N-cadherin和Vimentin等蛋白表达水平。利用在线miRNA靶基因预测网站对miR-151-3p下游靶基因进行预测,并结合PPI分析筛选关键靶基因。结果eEMS组(0.79±0.21)和nEMS组(0.62±0.15)miR-151-3p表达水平均显著低于对照组(1.02±0.17),差异有统计学意义(P<0.05),其中nEMS组最低。miR-151-3p过表达可显著上调ESCs细胞中miR-151-3p表达水平,抑制细胞增殖及迁移和侵袭能力,并下调MMP-2、MMP-9、N-cadherin和Vimentin等蛋白表达水平,上调E-cadherin蛋白表达水平(P<0.05);而抑制miR-151-3p表达显著降低ESCs细胞中miR-151-3p表达水平,促进细胞增殖、迁移和侵袭,并上调MMP-2、MMP-9、N-cadherin和Vimentin等蛋白表达水平,降低E-cadherin蛋白表达水平(P<0.05)。通过数据库预测出miR-151-3p下游34个交集靶基因,结合PPI分析筛选出4个关键靶基因:RC3H1、AGO2、AGO3和FXR1。结论MiR-151-3p可能在EMS患者的nEMS组织中低表达,其过表达可抑制ESCs细胞的迁移和侵袭。
Objective To explore the expression level of miR-151-3p in endometrial tissues of patients with endometriosis(EMS)and its effect on migration and invasion of endometrial stromal cells(ESCs).Methods The eutopic endometrial tissue(eEMS group)and non-eutopic endometrial tissue(nEMS group)of 87 EMS patients were collected.Meanwhile,47 cases of normal endometrial tissue from patients with benign gynecological diseases such as uterine fibroids who underwent surgical treatment were collected as the control group.qRT PCR method was used to detect the expression level of miR-151-3p in endometrial tissue of patients in each group.Human ESCs cells were isolated,identified and cultured,then miR-151-3p mimics,mimics NC,miR-151-3p inhibitors and inhibitors NC were transfected into ESCs cells,respectively.qRT PCR was used to detect the expression level of miR-151-3p in each group of cells;CCK-8 was used to check the cell proliferation ability;Transwell was used to detect the cell invasion and migration ability;Western blot was used to detect the expression levels of MMP-2 and MMP-9 and epithelial mesenchymal transition(EMT)markers E cadherin,N cadherin and Vimentin in each group of cells.Online miRNA target gene prediction websites were used to predict the downstream target genes of miR-151-3p,then the PPI analysis was used to screen for key target genes.Results The expression levels of miR-151-3p in the eEMS(0.79±0.21)and nEMS(0.62±0.15)groups were lower than those in the control group(1.02±0.17),with statistical significance(P<0.05)and the nEMS group was the lowest.Over-expression of miR-151-3p significantly up-regulated the expression level of miR-151-3p in ESCs cells,inhibited cell proliferation,migration and invasion,and down-regulated the protein expression levels of MMP-2,MMP-9,N cadherin and Vimentin,and up-regulated the protein expression of E cadherin level(P<0.05);while inhibiting the expression of miR-151-3p significantly reduced the expression level of miR-151-3p in ESCs cells,promoted cell proliferation,migrati
作者
谢震
赵健
吴娟
王雅丽
马丽霞
赵彩粉
XIE Zhen;ZHAO Jian;WU Juan;WANG Yali;MA Lixia;ZHAO Caifen(Second Department of Gynecology,Xingtai People′s Hospital,Xingtai 054000,Hebei,China;Third Department of Gynecology,Shijiazhuang People′s Hospital,Shijiazhuang 050011,Hebei,China;Cancer Examination Center of the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China)
出处
《医学研究与战创伤救治》
CAS
北大核心
2023年第5期456-462,共7页
Journal of Medical Research & Combat Trauma Care
基金
河北省医学科学研究课题计划(20210163)。
