MiR-329 Regulates The Expression Of CD146 In The Pathogenesis Of Endometriosis

Objective:1.To analysis the relationship between CD146,miR-329,MMP-9,VEGF and the development of endometriosis by detecting the expression of miR-329,CD146,MMP-9 and VEGF in endometriosis lesions and normal endometrial tissues.2.To investigate the potential molecular mechanism of CD146 and miR-329 in endometriosis by interfering with the expression of CD146 and miR-329 in normal endometrial stromal cells and ectopic endometrial stromal cells.Methods:1.Human normal endometrial tissues and ectopic endometrial tissues of patients with endometriosis were taken.CD146,MiR-329 and mmp-9 in endometrial tissues were determined by qRT-PCR,and the protein expression levels of mmp-9 and VEGF in endometrial tissues were determined by Western blotting.2.Normal endometrial stromal cells and ectopic endometrial stromal cells were extracted,isolated and purified,and the expression of CD146 in normal endometrial stromal cells and ectopic endometrial stromal cells was detected by flow cytometry.3.CD146 siRNA was transfected into endometrial stromal cells and ectopic endometrial stromal cells,and the expressions of CD146 and mmp-9 after transfection were determined by qRT-PCR.The protein expression levels of mmp-9 and VEGF were detected by Western blotting,so as to understand the effect of CD146 on the occurrence of endometriosis.4.Use riboFECT ? CP transfection reagent in ectopic endometrial stromal cells and normal endometrial stromal cells transfection miR-329 mimic and miR-329 inhibitor.The expression of CD146 after transfection was determined by qRT-PCR.Results:1.qRT-PCR detection revealed that the expression level of CD146 and mmp-9 in the ectopic lesion tissue was significantly higher than that in the normal endometrium tissue,and the expression level of mir-329 in the ectopic lesion tissue was significantly lower than that in the normal endometrium tissue.The expressions of mmp-9 and VEGF in the ectopic lesionswere significantly higher than those in the normal endometrial tissues,with statistically significant differences.2.The primary cells of normal endometrium and ectopic endometrium were successfully extracted and identified as endometrial stromal cells by purification,morphology and immunohistochemistry,which could be used for passage.Flow cytometry analysis showed that the expression ratio of CD146 in ectopic endometrium cells was higher than that in normal endometrium cells.3.Normal endometrial stromal cells and ectopic endometrial stromal cells were transfected with CD146 siRNA.The expressions of CD146 and mmp-9 were significantly down-regulated by qRT-PCR,and the differences were statistically significant.4.miR-329 inhibitor transfected normal endometrial stromal cells,and the expression level of CD146 in the transfection group detected by qRT-PCR was significantly higher than that of the non-transfection group.After transfection of ectopic endometrial stromal cells with mir-329 mimic,the expression of CD146 in the transfected group was significantly lower than that in the non-transfected group after qRT-PCR.The difference was statistically significant.Conclusion:1.In endometriosis,decreased the expression of miR-329,the expression of CD146 quantity increase in ectopic endometrium,miR-329 inhibits the expression of CD146,knocking the CD146 expression inhibition the expression of MMP-9 and VEGF,however,mmp-9 and VEGF as the typical representative of inflammation,attacks and new blood vessels form,it is concluded that miR-329 control CD146 expression to participate in the development of endometriosis.2.Further study on the specific mechanism by which CD146 or miR-329 or miR-329 regulates CD146 may lead to a better understanding of endometriosis,or it may be used as a drug treatment target to provide a new field of vision for the treatment of endometriosis.