Effects Of Endometrial Decidualization And Trophoblast Invasion On Pregnancy Outcome

Embryo implantation is an unique phenomenon to mammals inreproductive physiology, In the process of embryo implantation embryotrophoblast of matrix undergoes recognition, adhesion, invasion andendometrial decidual reaction as while, to establish tact and effectiveexchange between embryo trophoblast and matrix with Nutrients andinformation. It is the delicatecomplicated and multi-regulated consequencebetween embryo trophoblast and matrix.Any irregular step ofthe embryoimplantation may affect pregnancy outcomes. This study will follow thegene expression and regulation of trophoblast cell invasion ability bymethylation of E-cadherin gene promoter region and discusse theinfluence of Endometrial decidual and Sertoli cell invasion on the outcomeof pregnancy.PART ⅠGENE EXPRESSION PROFILE IN DECIDUA OFPATIENTS WITH RECURRENT SPONTANEOUSABORTIONObjectiveTo provide experimental basis for discovering possible mechanism ofRSA and specific markers screening, the abnormal gene expression in uterine decidua of patients with recurrent spontaneous abortion wasanalyzed by genome resorts.MethodsUterine decidua of patients with RSA and normal early pregnant werecollected and RNA was extracted for Microarray Hybridization and themicroarray data was analyzed by bioinformatics analysis. In addition, wevalidated the chip results with RT-PCR.Result1. Compared with control group,1656differentially expressed geneswere found in patient’s decidua with RSA, among which,1184up-regulatedgenes while472down-regulated genes were found.2. Differentially expressed genes involved in multiple biologicalprocesses and functions, including metabolism, cell-cell interaction,reproduction, development, external stimulus such as stress, cellproliferation and complement.3. Signal pathway analysis showed8signaling pathways mayinvolved in the progression of RSA, including FoxO family andAkt-mediated Class-I PI3K signaling pathways.4. Abnormal expression in microarray gene of patient’s decidua withRSA was verificated by semi-quantitative PCR. Compared with controlgroup, the expression of DKK1, GADD45A and GNLY genes wereincreased significantly while the DLK1expression was significantlyreduced, which indicated consistency with the results of gene chip.ConclusionGene expression profiling significantly changed in patient’s decidua with RSA is closely related with RSA. PART ⅡEFFECTS OF E-CADHERIN PROMOTERMETHYLATION ON INVASION ABLILITY OF HUMANTROPHOBLAST CELLSObjectiveTo investigate the role of adhesion molecules E-cadherin promotermethylation on the invasion ability of trophoblast cells in the embryoimplantation process.MethodsWe collected decidual-villi tissues, immunohistichemistry were usedto detected E-cadherin expression in different types of trophoblast cells. Tostudy the function of E-cadherin in trophoblast in vitro, we chosedHTR-8/SVneo cells and JEG-3cells for our study.5-aza-2’-deoxycytidine(5-Aza-cdR) was used to treat with the two cell lines. RT-PCR, western blotand Immunofluorescence were used to detected the E-cadherin expressionalternation compared with control group. we used MTT assay and flowcytometry to determine the optimum concentration of5-aza-2’-deoxycytidine (5-Aza-cdR) treating to HTR-8/Svneo cells; thendetected the cell invasion and migration by Transwell chambers. Results1. Immunohistochemistry detection of Chorionic villi-decidual tissueshowed: E-cadherin expression of invading the decidua chorion trophoblastcells (EVT) was decreased obviously compared to CTB (trophoblasts).2. RT-PCR western blot and Immunofluorescence detected that:E-cadherin gene expression was low in HTR-8/Svneo cells but highexpression in JEG-3cells. After the treatment with the demethylating agent5-aza-2’-deoxycytidine (5-Aza-cdR), E-cadherin expression had obviouslyrestored in the E-cadherin negative human intermediate trophoblastic cellline HTR-8/SVneo cells(p<0.01) but almost hadn’t changed in JEG-3cells.3. MTT assay and flow cytometry to determine the optimumconcentration of5-aza-2’-deoxycytidine (5-Aza-cdR) treating toHTR-8/Svneo cells:1μM and2.5μM5-aza-cdr had no effect on cellproliferation and apoptosis for HTR-8/SVneo cells.4. The motility and invasiveness of HTR-8/SVneo cells, as comparedwith the control cells, was significantly reduced.(p<0.01).Conclusionour results indicate that E-cadherin demethylation-modulated restoreof gene expression in HTR-8/SVneo cells results in a contact-mediatedinhibition of motility and invasion and suggest that an important role forhypermethylation-modulated of E-cadherin down-regulation in theintermediate trophoblast during implantation.