| Objective:1.Analysis of endometriosis glandular epithelial cell nuclear factor kappa NF-Bp65 regulating the expression of MMP9 in OPN.2.To investigate the changes of cell invasion of Bp65 cells in the endometrium of endometriosis with NF-kappa B before and after interventionMethods1.Collection of EMs in secretory phase eutopic endometrium in 12 cases,EMs gland epithelial cells in primary culture,purification,separation and determination of OPN by immunohistochemical method,the expression of NF-kappa Bp65 and MMP9 in the gland epithelial cells;respectively by gene silencing and plasmid,on patients with endometriosis at secretory phase endometrial epithelial cells in NF-kappa Bp65 intervention,Real Time-PCR and Western-blot detection of NF-kappa Bp65,OPN,mRNA and MMP9 protein expression.2.By invasion assay determination of invasive cells before and after intervention of NF-kappa Bp65 gene.Results1.Immunocytochemical results showed that NF-kappa Bp65,OPN and MMP9 were mainly located in the EMs gland epithelial cells in the cytoplasm.2.NF-kappa Bp65 after siRNA gene silencing in the gland epithelial cells of NFkappa Bp65,expression of MMP9 mRNA and protein significantly decreased,the difference was statistically significant(P <0.05),NF-Bp65 and MMP9 kappa mRNA and protein expression were significantly increased in NF-kappaB p65 plasmid,the difference was statistically significant(P<0.05)silencing siRNA gene into plasmid Bp65 kappa OPN expression did not change significantly,the difference was not statistically significant(P >0.05)3.In eutopic endometrial glandular epithelial cells,NF-kappaB p65 siRNA after the intervention of invasive cells were decreased compared with before intervention,significantly increased the cell invasion of NF-kappa Bp65 plasmid after the intervention,the difference was statistically significant(all P <0.05).Conclusion1.NF-kappa Bp65 siRNA silencing EMs eutopic endometrial epithelial cells,the expression of NF-kappa Bp65 and MMP9 decreased significantly;NF-kappa Bp65 was upregulated EMs eutopic endometrial epithelial cells,the expression of NF-kappa Bp65 and MMP9 increased significantly,NF-kappa B revealed abnormal expression of OPN is the key intervention regulation of MMP9 in endometriosis the patients in endometrium.2.NF-kappa Bp65 gene silencing EMs eutopic endometrial glandular epithelial cells,invasive epithelial cells decreased significantly;NF-kappa Bp65 plasmid EMs were up-regulated in eutopic endometrial glandular epithelial cells,invasive epithelial cells significantly increased,suggesting that in endometriosis NF-kappa Bp65 expression cell invasion by regulating OPN/MMP9. |
