Regulation Of Blastocyst Implantation By β1,4-galactosy Ltransferase I EGF, HB-EGF And TGF-α

Objective:To detect the roles ofβ1,4-galactosyltransferaseⅠEGF, HB-EGF and TGF-αduring the human implantation, their relationships, in order to investigate the control mechanism on embryo implantation.Methods:The RL95-2cells which imitate endometrium were transfected by vecters of HA-GalTⅠ, siGalTⅠ, pCDNA3.0, control siGalT. Incubation RL95-2cells with EGF, TGF-α, EGF and TGF-αantibodies separa-tely. RT-PCR, western-blot, Immunohistochemistry and fluorescence techni-ques were used to measure the level of EGF, HB-EGF and TGF-αin the transfected cells, and the level ofβ1,4-galactosyltransferaseⅠin the co-culture cells. Observing the effects ofβ1,4-galactosyltransferaseⅠ, EGF and TGF-a by RL95-2cells and JARcells which imitate embryo in the implantation.Results:1. After transfecting the cells for 60 hours, green fluorescent protein were detectede, Comparing with normal culture group and empty vector group, the synthesis ofβ1,4-galactosyltransferaseⅠof overexpressed group was dramatically, but declined in interference group,which telled the successful transfection. Results suggest the gene and protein level of EGF, HB-EGF and TGF-αincreased markedly in theβ1,4-galactosy-ltransferaseⅠoverexpressed group, the rate of embryo attachment (93.7±0.2%) was higher than normal culture group(85.3±0.5%), empty vector group (81.0±0.4%)and interference control group(85.7±0.6%), (p＜0.01), but declined in the interference group,and the rate of embryo attachment (66.9±0.3%) was lowest.(p＜0.01).2. After incubating RL95-2cells with EGF and TGF-a for 24h, Comparing with normal culture group, cyto-chemical staining showed that the synthesis of GAlβ1-4GlcNAc increased in treating EGF and TGF-a group. Andβ1,4-GalTⅠgene,β1,4-GalTⅠexpression increased, accodingly the rate of embryo attachment were(91.3±0.5%,91.5±0.4%), quitely higher than normal culture group (80.6±0.4%), while GAlβ1-4GlcNAc of the EGF and TGF-a antibodies treated group became lower,β1,4-GalTⅠgene,01,4-GalTⅠexpression decreased, their embryo attachment rate were67.6±0.2%,61.5±0.3%, and the rate of attachment was lower than the normal culture group80.6±0.4%, (p＜0.01).Conclusion:β1,4-GalTⅠinteracts with EGF,HB-EGF,TGF-αexpre-ssion in RL95-2cells, the expression ofβ1,4-GalTⅠis related to the embryo implantation. Results suggest regulating EGF, HB-EGF, TGF-αlevel may have effect on embryo identification and attachment. And EGF, HB-EGF, TGF-αact as a stimulus to embryo implantation,which may has relations with the expression ofβ1,4-GalTⅠin the endometrial cells.