| Mammalian embryo implantation is a complicated development process, itcontains the uterus and embryo between a series of synchronous coordinate change. Theprocess begins with identification of free of blastocysts and endometrial epithelial cells,location and adhesion, and then penetration into the uterus substrate. The processdepends on the establishment of endometrial accept state and the blastocyst invasivesuccessful nidation. Research shows that endometrial cells secrete a large number ofadhesion molecules, proteolytic enzym cytokines and related factor expression canpromote embryonic and endometrial mutual recognition and adhesion, and along withthe process of implantation.Objective: The change of β1,4-GalT-I expression level in endometrial cancercells (RL95-2cells),which can be seen as implantation period endometrial cells, as wellas the adhesive ability of embryos and endometrial was discussed after the treatment ofHCG.At the same time, EGFR pathways have been investigated. Control RL95-2cellsβ1,4-GalTI expression,testing the change of the corresponding matrix metalloproteinaseMMPs and extracellular matrix ECM expression, analysis and utilization of endometrialadhesion of influence, furthermoreβ1,4-GalT-I embryonic implantation and themolecular mechanism were discussed.Methods: Building model in vitro implantation in villi cancer cells (JAR cells) inorder to simulate before implantation embryo to endometrial cancer cells (RL95-2cells)in order to simulate before implantation endometrial; HCG β1,4-GalTI expressionlevel was detected by western blot, RT–PCR after the endometrial environment HCGlevel was upregulated or down regulated; galactosyl change was detected after theaction of HCG β1,4-GalT-I by Immune cell chemical dyeing; Gal β1,4-GlcNAcglycoprotein branch was checked by RCA-1lectin-blot after the action of HCG; Theprotein in EGFR pathway and the adhesive ability between RL95-2and JAR cells were detected after the treatment by western-blot,the fluorescence microscope and flowcytometry analysis,respectively.β1,4-GalT-I gene regulation plasmid (overexpression plasmid, interferenceplasmid), contrast interference plasmid and empty plasmid were transfected to RL95-2cell,respectively, RT-PCR and western blotting, immune cell chemical dyeing,immunofluorescence were used to check the expression level of β1,4-GalT-I,MMPs,as well as ECM after transfection. the adhesive ability between RL95-2and JARcells were detected by the fluorescence microscope and flow cytometry analysis invitro implantation model; Effect of HCG onβ1,4-GalT-I and MMPs-TIMP-ECMpathway was detected by RT-PCR.Results:(1) HCG is dose, time-effect β1,4-GalT-I expression in endometrialcells. The best condition is5x10-4ug/ul,36h (P <0.01);(2) EGFR, the nf-κ B, ICAM-1, EGF expression were obviously raised under the action of HCG (P <0.01);(3)HCG can promote adhesion effect of JAR cell to RL95-2cell (P <0.01);(4)Endometrial cells transfected by overexpression plasmid ofβ1,4-GalT-I gene groupcan obviously increase MMP2, MMP9, and significantly reduce LN, TIMP–1;interference group can obviously reduce MMP2, MMP9expression level, whileLN,TIMP-1expression level were significantly downregulated；interference group canobviously downregulate MMP2, MMP9while LN, TIMP-1were upregulated.(P＜0.01)；（5） over expressionβ1,4-GalT-I gene group can obviously improve theadhesive ability between JAR cell and RL95-2cell, while the the adhesive abilitybetween JARcell and RL95-2cell was decreased in interference group (P <0.01);(6)RT-PCR results show that obvious increase HCG MMP2, MMP9expression, decreaseLN, TIMP-1expression (P <0.01).Conclusion:(1)can regulate the expression of β1,4-GalT-I in endometrial cells(appear time, dose dependency relationship), promote adhesion ability of the JAR cellto RL95-2cell;(2) HCG regulate endometrial cells EGFR signaling pathway relatedmolecular expression to regulate and control the embryo implantation.(3) regulation ofβ1,4-GalT-I express influence embryo implantation may through the MMPs, ECMrelated signal pathways, regulate JAR cell to RL95-2cell adhesion Ability.(4) HCGregulate endometrial cells MMPs-TIMP-ECM path expression to regulate and controlthe embryo implantation. |
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