17-β Estradiol And Progesterone Regulate OPN Production And Cell Invasion In Endometriosis

Objective:1.Study the effect of 17-βestradiol(E2)and progesterone on the control of endometrium glandular epithelium(GE)cells’ osteopontin(OPN)and matrix metalloproteinase-9(MMP-9)metabolism in patients with endometriosis.2.Investigate the effect of 17-βestradiol(E2)and progesterone on the control of GE cells invasion in patients with endometriosis.3.Study the effect of siRNA-OPN on the control of GE cells’ OPN and MMP-9metabolism and cells invasion in patients with endometriosis.Methods1.Using immunohistochemistry to compare the expression and correlation of OPN and MMP-9 in the eutopic and ectopic endometrium of patients with endometriosis and in the normal endometrium.;2.We performed the primary culture of EECs and investigated the expression of OPN and MMP-9 in EECs regulated by 17beta-Estradiol(E2)and progesterone(P).OPN-specific siRNA interference was used to down-regulate OPN and to explore the change of MMP-9expression.Real-time RT-PCR,western blot analysis were used to detect the expression levels of OPN and MMP-9.Gelatin zymography was performed to observe the enzymatic activity of MMP-9 in the conditioned media.Transwell and wound scratch assays were carried out to investigate the migration ability of EECs.Results1.The expression levels of OPN and MMP-9 in normal endometrium were inferior to those in endometrium from patients with endometriosis;there is correlation between the expression levels of OPN and MMP-9;2.Treatment with 17-β estradiol or 17-β estradiol and progesterone significantly increased OPN and MMP-9 production in primary GE cells and GE cell s’ invasion compared with the control group(P<0.05).3.The expression levels of OPN and MMP-9 were remarkably decreased by siRNA interference in EEECs.Active MMP-9 was expressed less by siRNA treatment in EEECs.Conclusion1.OPN and MMP-9 was expressed more in endmetrium from patients with endometriosis;OPN may regulate the expression of MMP-9.2.The present study suggests that the up-regulation of MMP-9 via activation of OPN induced by estrogen may correlate with the migration of endometrial epithelial cells in endometriosis patients.