MiRNA-223 Expression In Endometrial Stromal Cells And Its Effect On Epithelial-to-mesenchymal Transition In Endometriosis

Part I Expression and clinical significance of HIF-1 a in serum of patients with endometriosisObjective:To study the expression level and diagnostic value of hypoxia inducible factor-1 α(HIF-1 α)in serum of patients with endometriosis(EMs).Through the study of serum level,the following problems were discussed:1.The expression of HIF-1 α in serum of patients with EMS,compared with the control group.2.The expression of HIF-1 α in serum of severe and mild EMS patients.3.Whether the expression of HIF-1 α in serum is different between endometrial proliferative phase and secretory phase.4.The receiver operating characteristic(ROC)curve and area under the curve(AUC)were used to diagnose EMs with HIF-1 α.The possible mechanism of HIF-1 α in the development of EMS was also discussed.Methods:the serum HIF-1 α levels of 36 patients with EMs(EMs group)and 43 patients with hysteromyoma(control group)were measured and analyzed by ELISA.The patients with clinical stage Ⅰ and Ⅱ in EMs group were mild EMS group,and the patients with stage Ⅲ and Ⅳ were severe EMS group.Receiver operating characteristic(ROC)curve and area under curve(AUC)were used to reflect the diagnostic level of HIF-1 α.Results:the expression of HIF-1 α in serum of EMS group was significantly higher than that of control group(P<0.001).The ROC curve and AUC results showed that HIF-1 α had a certain diagnostic effect on EMs,There was no significant difference in the expression of serum HIF-1 α between proliferative phase and secretory phase(P=0.593).The level of serum HIF-1 α in severe EMS group was significantly higher than that in mild EMS group(P<0.001).Conclusion:the expression level of HIF-1 a in serum of EMs patients is significantly increased,which can be used as a potential clinical diagnostic index.Part Ⅱ Expression of miRNA-223 in stromal cells of endometriosis and its effect on epithelial mesenchymal transitionObjective:To investigate the expression and clinical significance of miRNA-223 in endometrial stromal cells of EMs and its effect on epithelial mesenchymal transition of eutopic and ectopic endometrial stromal cells.The expression of miRNA-223 in eutopic and ectopic endometrial stromal cells of EMs,compared with the control group.2.The effect of miRNA-223 on epithelial mesenchymal transition in eutopic and ectopic endometrium of EMS.3.Effect of miRNA-223 on migration,invasion,proliferation and apoptosis of EMs eutopic and ectopic endometrial stromal cells.The possible mechanism of miRNA-223 in the development of EMs was also discussed.Methods:the endometrium of 26 patients with EMs(16 cases in EuSCs group,10 cases in EcSCs group)and 14 patients with uterine leiomyoma(control group)were collected.Primary endometrial stromal cells were isolated and cultured.The expression of miRNA-223 was detected by real-time quantitative polymerase chain reaction(qRT-PCR).Lentivirus and blank virus were transfected to obtain sh-miRNA-Western blotting was used to detect the expression of EMT related molecules N-cadherin,vimentin and Slug.Wound healing test and CCK8 method were used to detect cell migration and proliferation,and flow cytometry was used to detect cell apoptosis.Results:the expression of miRNA-223 in EMs group was significantly lower than that in control group(P<0.05).The expression of epithelial mesenchymal transition related molecules(N-cadherin,vimentin,slug)in sh-miRNA-223 group was significantly lower than that in sh-NC group(P<0.05).Compared with sh-NC group,the migration and invasion ability,proliferation and apoptosis rate of sh-miRNA-223 group were decreased(P<0.05).Conclusion:the expression of miRNA-223 in endometrial stromal cells is down regulated.Up regulation of miRNA-223 can reverse the EMT process and reduce the invasion,migration,proliferation and apoptosis of endometrial stromal cells.