儿童先天性胫骨假关节病变骨膜组织的蛋白质组学研究

Proteomics investigation on periosteum of children with tibia congenital pseudarthrosis

目的探讨基于串联质谱标签(Tandem Mass Tags,TMT)的蛋白质组学技术鉴定和分析先天性胫骨假关节(congenital pseudarthrosis of the tibia,CPT)病变骨膜组织中的差异表达蛋白。方法取CPT伴神经纤维瘤病1型(neurofibromatosis type 1,NF1)患者(NF1-CPT组)、CPT不伴NF1患者(非NF1-CPT组)和开放性胫骨骨折患者(对照组)的胫骨骨膜标本。采用基于TMT的蛋白质组学技术,对各组胫骨病变骨膜组织中差异表达的蛋白质进行鉴定和分析。设定倍数变化≥1.5或≤0.66且P值<0.05作为筛选差异表达蛋白的阈值。以在线工具DAVID和STRING生物信息学资源用于生成差异蛋白的GO注释、KEGG(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集和蛋白质-蛋白质相互作用(protein-protein interaction,PPI)。结果NF1-CPT组中有347种差异表达的蛋白质,其中212种上调、135种下调;非NF1-CPT组中有467种差异蛋白,其中281种上调、186种下调。NF1-CPT组与非NF1-CPT组共有差异蛋白231种,除HLA-DRB1(在NF1-CPT组中上调而在非NF1-CPT组中下调)外,其余230种差异蛋白在两组中表达趋势相同(上调117种、下调113种)。有116种蛋白质仅在NF1-CPT组中发生改变(上调94种、下调22种),236种蛋白质仅在非NF1-CPT组中改变(上调164种、下调72种)。与非NF1-CPT相比,NF1-CPT组的47种蛋白质变化1.5倍(P<0.05)。结论CPT病变骨膜组织与正常胫骨骨膜组织蛋白表达存在差异,NF1-CPT与非NF1-CPT的病变骨膜蛋白表达也存在差异,有助于对CPT发病机制的理解。

Objective To identify and analyze different proteins expression in the periosteum of congenital pseudarthrosis of the tibia(CPT)using tandem mass tags(TMT)proteomics.Methods The samples were divided into three groups,namely CPT with neurofibromatosis type 1(NF1)group(NF1-CPT group),CPT without NF1 group(nonNF1-CPT group)and control group(patients with open tibial fracture).A fold change≥1.5 or≤0.66 and P-value<0.05 was regarded as the threshold to screen differentially expressed proteins(DEPs).Subsequently,bioinformatics resources such as online tools DAVID and STRING were used to conduct GO annotation,KEGG pathways enrichment and protein-protein interaction(PPI)network with DEPs.Results A total of 347 proteins differentially expressed in NF1-CPT group,212 of which were up-regulated and 135 down-regulated.We identified 467 DEPs in nonNF1-CPT group,including 281 up-regulated and 186 down-regulated.Among of them,NF1-CPT group and nonNF1-CPT group shared 231 DEPs,except for HLA-DRB1 which increased in NF1-CPT group but decreased in nonNF1-CPT group.The remaining 230 DEPs showed the same expression trend in the two positive groups,including 117 up-regulated and 113 down-regulated.In particular,a total of 116 proteins were altered only in NF1-CPT group,including 94 up-regulated and 22 down-regulated.However,there were 236 proteins altered only in nonNF1-CPT group,including 164 up-regulated and 72 down-regulated.The results indicated that the pathogenesis of NF1-CPT was similar as nonNF1-CPT largely with a few differences.Finally,compared with nonNF1-CPT,there were 47 proteins changed 1.5-fold and P-value<0.05 in NF1-CPT group.Conclusion The proteins expression in the periosteum of CPT is different from that of normal tibia.The expression of periosteal protein is also different between NF1-CPT and nonNF1-CPT.The present study will deepen our understanding of the pathogenesis of CPT in the protein level.