DNA Microarray Analysis of Gene Expression in Eutopic Endometrium from Patients with Endometriosis

Pathogenesis of the endometriosis is complex and the etiology is still unclear. The objective of this study was to examine that endometrial gene expression in late secretory phase endometrium differs between patients with and without endometriosis. Five patients with proven advanced-stage endometriosis and 5 controls underwent endometrial biopsy in the late secretory phase. Analysis of eutopic endometrial gene expression was performed using Affymetrix gene arrays and differentially expressed genes were assigned to gene ontology groups based on overrepresented analysis using Database for Annotation, Visualization, and Integrated Discovery software. Four hundred sixty two genes were identified as up-regulated such as matrix metalloproteinase 10, cytochrome P450 family 24 subfamily A polypeptide 1, matrix metalloproteinase 3, chemokine (C-C motif) ligand 20, Rho family GTPase 1, interleukin 1-beta, and insulin-like growth factor binding protein 1. Six hundred forty three genes were down-regulated in all endometriotic samples. A lot of genes related with metabolic process, cellular ketone metabolic process and ncRNA metabolic processing were included. Expression patterns of selected five genes were validated by quantitative real time PCR. The results of this analysis support that the eutopic endometrium from patients with advanced-stage endometriosis has distinct gene expression profile from eutopic endometrium of control without endometriosis.

TNF system in eutopic endometrium from women with endometriosis

The role of the tumor necrosis factor (TNF) system in endometriosis is not completely clear and therefore was the focus of this study. In eutopic endometria obtained from women with (n = 41) and without (controls;n = 34) endometriosis during laparoscopy, the subcellular location and the percentages for TNF, TNFR1, TNFR2 and CD45 positive-cells were determined (immunohistochemistry);the protein concentration (ELISA) of the soluble receptors (sTNFR1 and sTNFR2) were measured in 4h-explants culture media and the TNF concentration was performed in ex vivo endometrial homogenates. The TNF, TNFR1 and TNFR2 mRNAs were analyzed by real-time PCR. In control endometria, TNF, TNFR1 and sTNFR1 proteins increased during the late secretory phase. In endometriosis endometria, the protein highest level of TNFR1 was reached during the early and the mid secretory phases and of sTNFR1 concentration during the proliferative phase;however, during the late secretory phase, both TNFR1 and sTNFR1 protein levels were reduced compared to the control endometria (p sTNFR2 concentrations, and TNF and TNFR2 mRNAs were similar in control and endometriosis endometria. Although TNFR1 mRNA was highly expressed, no significant differences were found between control and endometriosis endometria. In summary, this study establishes that TNF and TNFR1 protein expressions and the sTNFR concentrations in eutopic endometria from women with and without endometriosis are dependent on the menstrual cycle. The differences on the TNFR1 and sTNFR1 protein pattern between both groups, mainly during the mid and late secretory phases may play a role in the lower implantation rates observed in women with endometriosis, and also could be related to the altered cell death, which favor the augmented cell viability facilitating the characteristic endometrial implantation and growth outside the uterus in this disease.

Nerve Fibres Detection in Paired Eutopic and Ectopic Endometria from Women with Endometriosis: Correlation with Nerve Growth Factor Expression

Endometriosis, disorder characterized by the presence of endometrium outside the uterus cavity, is associated with chronic pelvic pain. Nerve growth factor (NGF) participates in development, repair and survival of neurons. We evaluated NGF and nerve fibres in eutopic and ectopic endometria from women with endometriosis and in normal endometrium from control patients without endometriosis. In endometrium collected during surgery for endometriosis (paired eutopic and ectopic endometria, proliferative n = 6 and secretory n = 7 phases) or during tubal ligation or hysterectomy for no endometrial disease (control, proliferative n = 6 and secretory n = 6 phases), NGF and neurofilament (NF) of nerve fibres were studied by immunohistochemistry measuring integrated optical density (IOD). Cytoplasmic NGF was detected in glands and stroma in all control, eutopic and ectopic endometrial samples;no statistical differences were found throughout the menstrual cycle. However, total (gland plus stroma) NGF IOD was higher in eutopic than in ectopic endometria during the secretory phase. NF was detected only in ectopic endometrium being eutopic and control endometria negative to this antibody, and no differences were observed between proliferative and secretory phases. Negative and significant correlation was found between NGF and NF expression in ectopic endometria only during the secretory phase. In conclusion, our results show a significant negative correlation between NGF and myelinated nerve fibres in secretory ectopic endometria. NGF expression was not significantly modified throughout the menstrual cycle in control and in endometriosis, even higher in eutopic than in ectopic tissues;the NGF secretion from lesions to pelvic cavity cannot be ruled out, and could explain the negative correlation observed. Although NF and NGF are not useful as diagnostic markers for endometriosis, their expression may be related to endometrial physiology and pathophysiology of the disease.

Increased expression of stathmin in eutopic endometrium of patients with endometriosis

Background Stathmin was identified as an endometriosis-related protein by comparative proteomics in our previous study.As a microtubule-destablizing factor, stathmin was shown to participate in the relay and integration of diverse intracellular signaling pathways involved in cell proliferation, differentiation, and many other cellular activities.To investigate whether stathmin is involved in the pathogenesis of endometriosis, we examined the expression of stathmin in eutopic endometrium of women with or without endometriosis.Methods Eutopic endometrium samples were collected from thirty-six patients who were diagnosed as endometriosis and the nineteen age-matched patients who were confirmed to be free of endometriosis surgically and histologically.The expression of stathmin mRNA was detected by real-time PCR, and its protein was detected by Western blotting and immunohistochemistry.Results Stathmin was overexpressed in eutopic endometrium of women with endometriosis detected by real-time PCR in mRNA levels and by Western blotting in protein levels, without significant difference between proliferative and 0secretory phase.Immunohistochemistry showed that stathmin protein was localized in both endometrial glandular and stromal cells throughout the menstrual cycle.Conclusions Stathmin is overexpressed in endometrium of patients with endometriosis and may play a role in the pathogenesis of endometriosis.

Ultrastructural Study of the Epithelium of Endometrial Glands in Women with Endometriosis:a Comparison with the Normal

ve The ultrastructure of endometrium women suffering from endometriosis was compared with those from women of normal reproductive age.