摘要
目的:基于蛋白组学观察尿道下裂患儿的差异蛋白特点,并分析相关蛋白ZEB1的表达。方法:收集我院2018年6月—2019年12月泌尿外科44例尿道下裂患儿外周血标本,同时收集44例健康体检儿童血液样本。取两组患儿血浆样本,提取总蛋白,进行二位凝胶电泳分离和图像分析,筛选出差异蛋白,利用质谱技术进行鉴定,并运用RT-PCR方法测定相关蛋白ZEB1mRNA的表达。结果:共筛选出44个差异表达蛋白,通过质谱分析和数据库检索论证,其中上调32个蛋白,下调12个蛋白;上调前5的蛋白有转录因子HES-1、膜联蛋白A3、E盒结合锌指蛋白1、锌指蛋白12和蓬乱蛋白,下调前5的蛋白则有锌转运蛋白10、肌球蛋白9、重组人碱性成纤维细胞生长因子受体2、胰岛素样生长因子2及肌球蛋白12。GO功能分析显示,差异蛋白主要参与了碱基化合物合成、细胞代谢、DNA转录、RNA代谢等生物学过程。KEGG通路分析表明这些蛋白主要集中在信号通路Th17细胞、Jak-STAT信号通路、Wnt信号通路等。通过STRING数据库构建差异蛋白相互作网络分析发现,差异蛋白ZEB1与STAT3、STAT6、CDH1、EB2、CDH2、CTBP1、SUMO1和TP53BP1关系较为密切。尿道下裂患儿血浆中ZEB1 mRNA表达量为(1.04±0.14),显著高于健康对照组,差异均有统计学意义(P<0.05)。结论:运用蛋白组学方法明确了尿道下裂患儿血清差异蛋白质的表达,ZEB1在尿道下裂发生过程中具有重要作用。
Objective:To investigate characteristics of differential protein in children with hypospadias based on proteomics and analyze related protein ZEB1 expression.Methods:From June 2018 to December 2019,44 children with hypospadias were selected and the peripheral blood samples were collected.Meanwhile,the blood samples of 44 children with health check were also obtained.The serum samples of children in two groups were attained and the total proteins were extracted.Two gel electrophoresis separation and image analysis were used to select differential proteins.Then,mass spectrometry was employed to identify,and RT-PCR method was applied to determine the expression of related protein ZEB1 mRNA.Results:Forty-four differentially expressed proteins were screened by mass spectrometry and database retrieval,which 32 proteins were up-regulated,and 12 proteins were down-regulated.The first five proteins with up-regulation were the transcription factor HES-1,membrane protein A3,zinc finger E-box binding homeobox 1,zinc finger protein 12 and DVL.While,the first five proteins with down-regulation were zinc transporters 10,myosin 9,human recombinant basic fibroblast growth factor receptor 2,insulin-like growth factor 2 and myosin 12.GO function analysis indicated that the differential proteins were mainly involved in the biological processes such as nucleobase-containing compound metabolic process,cell metabolism,DNA,RNA transcription metabolism process.KEGG pathway analysis showed that these proteins were mainly concentrated in the signal path Th17 cells,Jak-STAT signal pathway,Wnt signaling pathways,etc.The differential protein network built by STRING database indicated differential protein ZEB1 had close relationship with STAT3,STAT6,CDH1,EB2,CDH2,CTBP1,SUMO1 and TP53 BP1.Plasma ZEB1 mRNA expression in children with hypospadias was(1.04±0.14),which was significantly higher than that of healthy controls.The difference all presented statistically significant(P<0.05).Conclusion:The use of proteomics approach has been clear about
作者
农一敏
张宝欣
乌日尼勒图
NONG Yimin;ZHANG Baoxin;WURINILETU(Department of Pediatric Surgery,Shenzhen Baoan Women's and Children's Hospital,Shenzhen,Guangdong,518102,China)
出处
《临床泌尿外科杂志》
CAS
2021年第4期288-292,共5页
Journal of Clinical Urology
基金
深圳市宝安区科创局(No:2017JD013)。
