Effects Of Genome Gene Expression Profiling Of Progesterone And Interferon-tau On Bovine Endometrial Cell Cultured In Vitro

During the establishment of pregnancy of cow, interferon-tau (bIFN-τ) as the identification signal of conceptus and progesterone (P) as the maternal pregnancy hormone joint act in endometrium, which caused the changes of genes be related to pregnancy recognition and embryo implantation in endometrium, meanwhile the signal accomplished from bIFN-τleading to P dominant.In this study, we used the Agilent bovine genome commercial microarray to examine the differential gene expression profiling of in endometrium with the treatment of interferon-τ(bIFN-τ), progesterone (P) and interferon-τplus progesterone (bIFN-τ+P). The results of array analysis are as follows:(1 Successfully cultured cow endometrial cells in vitro model. Cultured endometrium cells in vitro could grow in DMEM-Ham's F12 medium which comprised 100 IU/mL Penicillin,100μg/mL streptomycin,2μmol/mL Glu and 20% fet al bovine serum. Cells type in primordial and passage generation were endothelial-like cells which were from endometrium endothelial cells, and fibroblast-like cells which from endometrium stroma-cell.(2)Difference expression, clustering analysis showed that:there are 1401 (579 up and 822 down) differential display genes in progesterone group contrast to control group respectively,2380 (1383 up and 977 down) differential display genes in interferon-τgroup contrast to control group respectively,2389 (1098 up and 1091 down) differential display genes in progesterone plus interferon-τgroup contrast to control group respectively,2308 (841 up and 1467 down) differential display genes in progesterone plus interferon-τgroup contrast to interferon-τgroup respectively,1360 (683 up and 677 down) differential display genes in progesterone plus interferon-τgroup contrast to progesterone group respectively.(3)Gene Ontology(GO) enrichment analysis showed that: in iprogesterone group contrast to control group, in interferon-τgroup contrast to control group, in progesterone plus interferon-τgroup contrast to control group, in progesterone plus interferon-τgroup contrast to interferon-τgroup, in progesterone plus interferon-τgroup contrast to progesterone group, we found 63,50,46,37and 32 significant (P<0.05 and 0.01) GO correlated to pregnancy establishment respectively, primary focusing on immune response and regulation of immunologic process, cell proliferation and differentiation, angiogenesis, cell adhesion, glucose and amino acid tansport, female pregnancy and embryo development, interaction of cytokine, hormonal synthesis, secretion and response to hormonal stimulus, proteolysis, DNA methylation and histone acetylation et al, suggested that progesterone and interferon-τeffect the expression of in pregnancy recognition and embryo implantation process.(4)Pathway enrichment analysis showed that:in iprogesterone group contrast to control group, in interferon-τgroup contrast to control group, in progesterone plus interferon-τgroup contrast to control group, in progesterone plus interferon-τgroup contrast to interferon-τgroup, in progesterone plus interferon-τgroup contrast to progesterone group, we found 11,14,15,13,12 significant (P＜0.05 and 0.01) pathway correlated to pregnancy establishment respectively, primary focusing on GnRH signaling pathway, ECM-receptor interaction, Cytokine-cytokine receptor interaction, MAPK signaling pathway and TGF-beta signaling pathway et al. suggested that progesterone and interferon-τmaybe effect the capital genes in these pathway and to regulate the pregnancy establishment.(5)During the construction of Co-Expression-Network, we found 8 Co-Expression genes between iprogesterone group contrast to control group and progesterone plus interferon-τgroup contrast to control group, they are CXCL2, CCL5, IL6, IL8, CSF2, FGF9, FGF1 and TNF, suggested that these genes may paly an important role in regnancy recognition and embryo implantation. (6)Reliability evaluation from microarray data showed that:the C.V range from 4.84%-12.67% within-arrays, lower the standard of within C.V 15%, meanwhile qRT-PCR was used to verify the microarray data, the results showed that the coefficient correlation between these two technology was:0.8739±0.06367, these resultes demonstrate that the microarray technique used in this study is accurate and reproducible.