The MiRNA Expression In Endometriosis And Its Relation To Pathogenesis

objective:To study expression and Clinical significance of Dicer and Drosha in Eutopic endometrial tissue and Ectopic endometrial tissue of endometriosisMethod:Use RT-PCR and western blot to detect expression difference of Dicer and Drosha in Eutopic endometrial tissue and Ectopic endometrial tissue; Groom Eutopic endometrial tissue by way of in virtro, interfere Dicer and Drosha with siRNA, then compare Proliferation difference of endometrial tissue between the non-interfered group and the interfered group, and use Flow cytometry to detect Apoptosis of cells; Lastly, ELISA detect expression of and use western blot to detect expression change of Apoptosis-suppression protein bcl2and Pro-apoptotic protein bax.Result:1. Expression of Dicer and Drosha in Ectopic endometrial tissue is lower than that of Eutopic endometrial tissue.2. Proliferation of Ectopic endometrial cell for Dicer and Drosha group interfered with siRNA is accelerated, while cell Apoptosis is decreased.3. The expression of Transforming Growth Factor-β1in Eutopic endometrial tissue of Dicer and Drosha group interfered with siRNA is increased; Expression of apoptosis suppression Protein bcl-2is increased, but that of Pro-apoptotic protein bax is decreased. Conclusion:Dicer and Drosha, the important regulator for miRNA can affect Proliferation and Apoptosis of cell by affecting expression of TGF-β1and bcl-2/bax, further contributing to formation of endometriosis. Objective:To make a research on expression difference of miRNA gene in Eutopic endometrial tissue and Ectopic endometrial tissue for endometriosis patients and dig into the pathogenesis of endometriosis on a molecule basis.Methods:Specimen are taken from Eutopic endometrial tissue and Ectopic endometrial tissue of endometriosis patients for MicroRNA chip scanning to identify difference MiRNA of the two, predict target genes for the difference MiRNA, make relevant statistics for miRNA frequency of regulating target genes, classify the target genes corresponding to difference miRNA on a function basis and analyze signaling pathways possibly participated by those target genes to configure Regulatory network of MicroRNA-Gene. Furthermore, Network biology and graph theory are employed for analysis so as to determine key miRNA and controlled target genes.Result:1. Through analysis of miRNA chip,12miRNAs with increasing expression (representative:hsa-miR-202) and39miRNAs with decreasing expression (representative:hsa-miR-33b) are selected based on taking Pval<0.05as effective gene, ratio>2as expression increasing standard and ratio<0.5as expression decreasing standard. 2. By predicting miRNA target gene of increasing difference and decreasing difference respectively through online software and then analyzing with Go function of difference target gene, it is found that target gene of miRNA with decreasing difference takes SMC1A as representative, which is enriched in such biological functions as cellular process, biological process and biological regulation, while target gene with increasing difference takes CACNA2D3as representative, which is enriched in such biological functions as cellular process, biological process and metabolic process.3. It is found in analyzing the pathways which are possible to be affected by target gene with difference miRNA that target gene of miRNA with decreasing difference are mainly concentrated in signaling pathways of OOCYTE MEIOSIS and Tumorigenesis, while target gene of miRNA with increasing difference are mainly concentrated in signaling pathways of MAPK and Tumorigenesis.4. Regulatory network diagram of miRNA and target gene are established for further study of difference miRNA function.Conclusion:The12miRNA (like hsa-miR-202&hsa-miR-508-3p) are increasingly expressed in ectopic endometrium, while the39miRNA (like hsa-miR-33b and hsa-miR-34b) are decreasingly expressed; Target genes regulated by these difference miRNA mainly participate in metabolic regulation and gene transcription process, which will further affect signaling pathways of MAPK, OOCYTE MEIOSIS and Tumorigenesis, thus leading to generation and development of endometriosis.