目的 分析单核细胞增生李斯特氏菌(简称单增李斯特菌)感染人绒毛膜滋养层细胞HTR-8/Svneo过程中引起的细胞蛋白质组变化.从宿主细胞分子水平对单增李斯特菌感染胎盘的机制进行研究.方法 运用TMT(Tandem Mass Tags)蛋白质组学技术对HTR-8/Svneo感染组和非感染组蛋白质组进行定量分析,运用GO(Gene Ontology)、KEGG(Kyoto Encyclopedia of Genes and Genomes)数据库对差异表达蛋白进行分子功能、生物过程及代谢途径富集分析.结果 共鉴定肽段76 285个,可定量蛋白6 979个.HTR-8/Svneo感染过程中356个蛋白发生了显著性差异表达.153个表达量上调,203个表达量下调.差异表达量倍数最高的蛋白为微管马达蛋白(B4DYE2),最低为翻译起始因子1(Q6IAV3).GO功能富集和KEGG代谢途径富集结果表明,上调蛋白参与的生物过程和代谢途径集中在微管、微丝运动,细胞自噬等方面,下调蛋白集中在碳代谢、氮代谢、氨基酸生物合成、核酸代谢等初级代谢途径和泛素介导的蛋白质水解等方面.结论 单增李斯特菌侵袭HTR-8/Svneo过程中,宿主细胞的基础代谢可能发生了显著降低,并处于较高的自噬水平.细胞迁移诱导透明质酸酶CEMIP(Q8WUJ3)的显著上调说明该蛋白可能在调节滋养层细胞迁移、侵袭能力进而造成不良妊娠方面具有重要作用,为单增李斯特菌感染胎盘分子机制研究提供新的思路.
Comparative proteomics analysis of human villous trophoblasts during the infection of Listeria monocytogenes
This study was aimed at analyzing the changes in the cell proteome of HTR-8/Svneo human villous trophoblasts during infection with Listeria monocytogenes,to explore the mechanisms involved in L.monocytogenes infection of the placenta at the molecular level in host cells.Tandem Mass Tag(TMT)proteomics was used to compare the quantitative proteome be-tween the HTR-8/Svneo infection group and control group.The Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases were used to analyze enrichment in the molecular functions,biological processes,and metabolic pathways of differentially expressed proteins.A total of 76 285 peptides and 6 979 quantifiable proteins were identified.A total of 356 proteins showed significant differential expression:153 up-regulated and 203 down-regulated.The protein with the high-est fold change was microtubule motor protein(B4DYE2),and the protein with the lowest fold change was translation initia-tion factor 1(Q6IAV3).GO functional enrichment and KEGG metabolic pathway enrichment analyses indicated that the up-regulated proteins were involved primarily in biological processes and metabolic pathways such as the movement of microtubules and microfilament as well as autophagy,whereas the down-regulated proteins were involved mainly in primary metabolic pathways,such as carbon metabolism,nitrogen metabolism,amino acid biosynthesis,and nucleic acid metabolism,as well as ubiquitin mediated protein hydrolysis.In the process of infection of HTR-8/Svneo by L.monocytogenes,the basal metabolism of host cells may be significantly diminished,and the autophagy level may be high.The significant up-regulation of cell migration-inducing and hyaluronan-binding protein CEMIP(Q8WUJ3)suggested an important role of this protein in regulating the migration and invasion of trophoblast cells,thus causing adverse pregnancy outcomes.Our findings may provide new ideas for the study of the molecular mechanism of L.monocytogenes infection of the placenta.
万方数据
维普
