ABSTRACT
Growth arrest specific protein 6 (GAS6) plays an important role in the occurrence and development of tumors, and its signal transduction is involved in cell proliferation, adhesion and migration, but its related functions and molecular mechanisms in endometriosis (EMs) are still unclear. In this study, we searched and downloaded the transcriptome datasets of EMs from GEO database and performed GEO online analysis, and then screened out the differentially expressed genes and performed cluster analysis based on GO and KEGG pathway. The mRNA levels of the differentially expressed genes shared by more than three datasets were verified by qRT-PCR in the endometrium of ten women with no endometriosis and no clear disease and the ectopic endometrium of 11 patients with ovarian chocolate cysts. Immunohistochemistry and qRT-PCR were used to verify the expression of GAS6 and epithelial mesenchymal transition (EMT) marker genes, and immunofluorescence was used to co-label GAS6 and E-cadherin in endometriosis clinical samples. In this study, a total of 47 differentially expressed genes were screened out of the four transcriptome datasets, which were mainly enriched in processes such as cell migration and related signal pathways such as MAPK, PI3K-AKT, and tight junction. The mRNA levels of the nine differentially expressed genes shared by more than three datasets in endometriosis patients were consistent with the results of bioinformatics analysis. GAS6 expression levels in ectopic endometrium of EMs patients are higher than the control group (P < 0. 05), and EMs patients have the characteristics of EMT in the ectopic endometrial tissue, that is, the expression of E-cadherin is down-regulated (P < 0. 05) and the expression of vimentin is up-regulated (P < 0. 01). The expression of E-cadherin in the ectopic endometrial glandular epithelial cells of EMs patients is low while the expression of GAS6 is up-regulated, suggesting that GAS6 may mediate the EMT process in endometriosis. In conclusion, this study reveals that GAS6 is highly expressed in endometriosis patients and may mediate the EMT process to participate in the occurrence and development of endometriosis, providing a potential target for clinical treatment of endometriosis.
ABSTRACT
Objective:To investigate Cx43 expression and gap junctional intercellular communication(GJIC) in ectopic and eutopic endometrial stromal cells in endometriosis(EMs),and to explore the influence of aberrant GJIC in stromal cells on pathogenesis of EMs. Methods:The stromal cells were isolated from samples including 24 ectopic endometriotic tissues located in ovaries,41 eutopic endometria with endometriosis and 30 normal endometria. The endometrial stromal cells models were established in vitro by being cultured in manual conditions mimicked with estrogen and progesterone. Laser scanning confocal microscopy(LSCM) was used to determine the expression of Cx43 protein and the function of GJIC in three groups stromal cells. Results:The success rate of isolation and culture of endometriotic stromal cells was 45.8%(11/24);of eutopic endometrial stromal cells with EMs was 92.7(38/41);of normal endometrial stromal cells was 93.3%(28/30). The purities of ectopic and eutopic endometrial stromal cells were 95% and 98% respectively. The level of Cx43 protein and the function of GJIC in stromal cells from ectopic endometrial tissues were much lower than those from the other two groups,the highest level of Cx43 protein and the function of GJIC were observed in normal endometrial stromal cells group,and the differences among these groups were significant (P﹤0.01). Conclusions:(1) It will be helpful to establish models of normal,ectopic and eutopic endometrial stromal cells in vitro simultaneously when investigating the pathgenesis of EMs. (2)Downregulation of Cx43 expression and aberrant function of GJIC are related to pathogenesis of EMs. Regulation of Cx43 or GJIC in endometrial stromal cells is implied to be a potential strategy to treat EMs.