The effect and mechanism of CXCR4 targeted by miR-30b on endometrial stromal cells in mice with adenomyosis
Objective:To explore the effects of miR-30b on endometrial stromal cells(ESCs)in mice with adenomyosis(AM)as well as its potential regulatory mechanism.Methods:A total of 24 ICR female mice were selected to make AM animal model,and 4 ICR mice were selected as normal controls.HE staining was used to observe the pathological changes of the uterus,and reverse transcription fluorescence quantification polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-30b in the uterus of AM model mice and normal mice.ESCs of AM model mice were isolated and cultured,and randomly divided into the control group,miR-30b overexpression group(miR-30b group)and negative control group with miR-30b overexpression(miR-NC group).The expression of miR-30b was detected by qRT-PCR,and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay,transwells assay and flow cytometry were used to detect cell proliferation,migration and apoptosis,respectively.Western blot was used to detect the expression levels of CXC chemokine receptor 4(CXCR4)and vascular endothelial growth factor(VEGF)in cells.The targeting relationship between miR-30b and CXCR4 was verified by dual luciferase report.Results:Among the 24 mice,22(91.67％)were successful in AM modeling.Compared with normal mice,the uterus of AM model mice was distorted and strong,with prominent adenomyopathy nodules on the surface,uterine myometrium development disorder,endometrial interstitial infiltration,and endometrial interstitial cells invaded the inner muscle layer.The expression of miR-30b was significantly lower in the uterine tissue of AM model mice(P＜0.05).while overexpression of miR-30b reduced the proliferation and invasion abilities of ESCs and accelerated cell apoptosis of model mice(P＜0.05).Furthermore,overexpression of miR-30b significantly decreased the expression levels of VEGF and CXCR4 protein in ESCs(P＜0.05).It was found that miR-30b directly targeted CXCR4.Conclusions:miR-30b may directly regulate CXCR4 expression,resulting in the promotion of proliferation and invasion of ESCs cells,inhibition of cell apoptosis,and involvement in the pathogenesis of AM.
万方数据
维普
