| The research background and purpose:Endometriosis(EMs)is one of the common benign gynecological diseases,the main manifestations are infertility and pelvic pain.Until now,the etiology and pathogenesis,which are still unclear,so constructed in vitro model of endometrial cells has become an inevitable trend in the researching field of obstetrics and gynecology,aiming to provide reliable platform for researching cells in vitro of endometriosis.Epithelial to mesenchymal transition(EMT)gives the characteristics of cell migration and invasion,which may be the prerequisite of established endometriosis.This process is regulated by signaling pathways and transcription factors,snail is one of the transcriptional factor,which may play important role.The more research of EMT and snail were expressed in tumors,recent studies show that EMT may be involved in endometriosis,whether it is mediated by the transcription factor snail is not related reports.In this study,we through culturing the primary cells of normal endometrial、eutopic endometrial in adenomyosis and ectopic endometrial in endometriosis,which can be detected the differences between them by cell morphology and can be measured the migration and invasion.To explore whether snail mediated epithelial-mesenchymal transition or not and the expression of the related markers molecules of EMT:E-cadherin、β-catenin and vimentin from protein levels to provide some basis of molecular theory for the pathogenesis of endometriosis.Methods:1.In vitro model: Though primary culture to establish the cells in vitro model of normal endometrial、eutopic endometrial in adenomyosis and ectopic endometrial in endometriosis and detect morphology of cells。2.Wound-healing assay: Though primary culture to compare the migrate ability of cells.3.Invasion assay: Though primary culture to compare the invasive ability of cells.4.Though methods of immunohistochemistry SP(streptavidin peroxidase)and western blot to explore the expression of transcription factor snail and the related markers molecules of EMT: E-cadherin、β-catenin and vimentin in different group.Results:1.The success rate of normal endometrial cells 、 eutopic endometrial in adenomyosis and ectopic endometrial in endometriosis were 91.43%(32/35),91.30%(21/23),89.47%(17/19),and they had similary cell morphology between ectopic endometrial cells and normal endometrial cells.2.Wound-healing assay results:The difference of channel width before and after24 hours of ectopic endometrial cells in endometriosis group and control group was statistically significant(P <0.05);The difference of channel width before and after 24 hours of ectopic endometrial cells in endometriosis group and eutopic endometrial in adenomyosis group was statistically significant(P <0.05).3.Invasion assay results: The cells number of penetration of control group were(39.36 ± 17.33);eutopic endometrial in adenomyosis group were(59.28 ± 11.91);ectopic endometrial in endometriosis group were(59.28 ± 11.91),eutopic endometrial in adenomyosis group and ectopic endometrial in endometriosis group were increased with control group(P<0.05).4.Immunohistochemical results: The positive ratio of E-cadherin in the group of EMs was 15.79%,the group of ectopic endometrial in adenomyosis was 30.43%,which were decreased with control group(P<0.05),The positive ratio of vimentin in the group of EMs was 89.47%,the group of ectopic endometrial in adenomyosis was86.96%,which were increased with control group(P<0.05);The positive ratio of β-catenin in the group of EMs was 42.11%,the group of ectopic endometrial in adenomyosis was 56.52%,which were decreased with control group(P<0.05);The positive ratio of snail in the group of EMs was78.95%,the group of ectopic endometrial in adenomyosis was 82.61%,which were increased with control group(P<0.05),The positive ratio of snail in the group of eutopic endometrial in adenomyosis was 56.52%,which was decreased with the group of ectopic endometrial in adenomyosis(P<0.05)。5.WB results: The expression of E-cadherin and β-catenin in the groups of EMs and AM were decreased with control group(P<0.05);The expression of vimentin in in the groups of EMs and AM were increased with control group(P<0.05);The expression of snail in the group of EMs was increased with the group of control and AM(P<0.05),The expression of snail in the group of ectopic endometrial in adenomyosis was increased with control group(P<0.05),The expression were no differences between the group of eutopic endometrial in adenomyosis and the control group(P>0.05).6.The Spearman analysis showed that significant correlations between Ecadherin and vimentin,E-cadherin and snail in the groups of EMs and ectopic endometrial in adenomyosis(P<0.05);There were no correlations in the control group(P>0.05)。Conclusion:1.The migration and invasion of ectopic endometriosis cells was significantly stronger than normal endometrial cells;But ectopic endometrial cells and normal endometrial cells had similarity cell morphology,suggested that ectopic endometrial cells may originate from normal endometrial cells.The endometrial cells may be adapted to the specific microenvironment after implanted,which could cause different types of pelvic endometriosis.2.The expressions of molecular markers of epithelial cells E-cadherin and β-catenin were reduced in the patients of endometriosis and adenomyosis while the expression of molecular marker of mesenchymal cell vimentin was increased and the expression of transcription factor snail was increased too,indicating that EMT may correlate with occurrence of endometriosis and the transcription factor snail may be involved regulation of the process,which was to say that EMT may regulated the migration and invasion in endometriosis by transcription factor snail to downregulate the expression of E-cadherin through specific pathway. |
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