Expression Of E4BP4 And COMT Genes Related To Implantation In Eutopic Endometrium From Patients With Endometriosis

PART ONEObjectiveThis study was designed to detect the expression and localization of E4BP4 in human eutopic endometrium from patients with endometriosis.Materials and MethodsProliferative endometrium and secretory endometrium were collected respectively from patients with endometriosis(n = 13) and the control(n = 13).Reverse Transcriptase PCR,Real-Time quantitative RT-PCR and Western blotting were performed to examine the relative level of E4BP4 mRNA and protein in eutopic endometrium,Immunohistochemistry were used to determine the E4BP4 protein localization in human endometrial tissues.ResultsE4BP4 mRNA and protein relative levels in patients with endomeosis were significantly decreased both in the secretory phase and proliferative phase compared with the control group;Immunohistochemistry showed that E4BP4 protein was expressed both in the endometrial stroma and endometrial glands.E4BP4 expression in the endometrial glands was detected both in cytoplasm and nuclei.In stroma,there was more intense E4BP4 immune staining in the nuclei compared with the cytoplasm. There was a significant difference between two groups in the secretory phase and no significant differences in the proliferative phase.ConclusionsE4BP4 might mediate in disturbing that embryo implantation in EM PART TWOObjectiveBy investigating the variations of COMT(catechol- O-methyltransferase) mRNA and protein during the mouse estrous cycle and in eutopic endometria of patients with EM and the regulations by estrogen and progesterone。The study tried to give novel explications for the function of COMT in the reproductive processes.MethodsRT-PCR,real-time quantitative RT-PCR and Western blotting were used to examine COMT mRNA and protein in mature and prepubertal mice uterine,ovarian tissue and eutopic endometria of patients with EM.Results1.COMT mRNA and protein of uteri and ovaries,were discovered highly expressed at estrous significantly,compared with proestrus,metaestrus,and diestrus in the mouse estrous cycle.2.Treatment with estradiol resulted in a decrease in COMT mRNA level in ovarian tissues of prepubertal mice.3.COMT mRNA of pregnant mouse uterus was down-regulated in reference to non-pregnant mouse uterus.4.There were no significant differences of COMT expression between EM and the control in the proliferative endometrium and the secretory endometrium.Conclusions1.The level of COMT mRNA in mouse uteri and ovaries fluctuates in the estrous cycle,with a peak at estrus.Estrogen,not progesterone,may play an important role in the down-regulation of the COMT mRNA level.2.There were no obvious relevance between COMT expression and the alteration of microenvironment of embryo implantation in patients with endometriosis. PART THREEObjectiveThe alteration of gene expression profiles in endometriosis was evaluated using Illumina BeadChip microarrays.MethodsPatients with endometriosis(n = 8) and non-endometriosis(n = 9) were selected for gene profile microarray analysis.Results422 DE(differential expression) genes were found with more than 2-fold or less than 0.5-fold changes in endometriosis group compared with non-endometriosis group, including 236 genes down-regulated and 186 up-regulated.The DE genes involved in cell signaling,cell adhesion and differentiation,immune adjustment and inflammatory response that occur in the implantation window.ConclusionsThe changed gene expression profiles in response to endometriosis can specify the deterioration in embryo implantation and provide gene targets for further researches.