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幽门螺杆菌感染相关胃癌的差异蛋白分析

Differential proteome profiling of Helicobacter pylori-associated gastric cancer

摘要:

目的 利用核素标记相对和绝对定量(iTRAQ)技术筛选幽门螺杆菌(Hp)感染相关胃癌的差异蛋白,以便寻找胃癌的生物学标志物和药物治疗靶标.方法 分别收集人胃癌细胞HGC-27及与Hp共培养24h的HGC-27的各组蛋白,iTRAQ试剂标记后利用强离子交换柱(SCX)分离,经串联质谱鉴定及相对定量;同时通过查阅Pubmed数据库中1995年1月至2014年7月公开发表的文献,其中使用任一种差异蛋白质组学方法鉴定Hp感染相关胃癌的差异蛋白;将iTRAQ数据与文献报道数据进行荟萃分析,对获得的差异蛋白采用PANTHER数据库进行基因功能聚类(GO)分析、PANTHER蛋白分类及路径(Pathway)分析.结果 通过iTRAQ标记鉴定出Hp感染相关胃癌细胞HGC-27的差异蛋白共47个,其中下调表达的37个,上调表达的10个.通过文献检索共纳入符合要求的文献7篇,其中2个以上实验室(包括iTRAQ实验)重复发现的蛋白为21个.将iTRAQ数据与文献报道数据荟萃分析得到65个差异蛋白,进行GO分析显示,生物学过程主要集中在细胞组分重组和各种代谢过程;分子功能主要集中在核酸、DNA结合,具有催化活性和转移酶活性;PANTHER蛋白分类主要为分子伴侣、结合蛋白、骨架蛋白、转氨酶和转录因子;Pathway分析显示差异蛋白主要参与帕金森疾病通路、凋亡信号通路和促性腺激素释放激素(GnRH)通路.结论 筛选出多种与Hp感染相关胃癌的差异蛋白,为胃癌患者的早期诊断提供可能的生物学标志物和药物治疗的靶标.

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abstracts:

Objective To screen for the differential expressed proteins in Helicobacter pylori-associated gastric cancer by using isobaric tags for relative and absolute quantitation (iTRAQ) labeling and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS),and to explore the potential targets for biomarkers and biological therapy.Methods Collection the proteins of gastric epithelial cells HGC-27 and HGC-27 co-cultured with Hp,then strong cation exchange (SCX) separation after iTRAQ reagents labeled,then identification and quantitative via tandem mass spectrometry (MS/MS).The literature about differential expressed protein in Hp-associated gastric cancer by any of proteomic techniques published between January 1995 and July 2014 were selected by searching PubMed database.The screening results of iTRAQ labeling and the reported data were meta-analyzed by PANTHER database.Results Forty-seven differential expressed proteins were identified by using iTRAQ labeling.Among the 47 proteins,37 were up-regulated and 10 were down-regulated.Seven pieces of eligible literature were obtained by information retrieval,21 were reported by two or more labs.The iTRAQ data and the reported data meta-analyzed to get 65 differential expressed proteins.The gene ontology (GO) analysis demonstrated the major molecular functions including nucleic acid and DNA binding,catalytic activity and transferase activity as well as the cellular component organization and metabolic process in biological processes.PANTHER Protein Class,mainly for chaperone,binding protein,cytoskeletal protein,transferase and transcription factor.The differential proteins were mainly involved in Parkinson disease pathway,apoptosis signaling pathway and gonadotropin releasing hormone receptor pathway.Conclusion Screening for a variety of differential expressed proteins in Hp-associated gastric cancer may provide the potential gastric cancer-related biomarkers and therapeutic targets in early diagnosis and treatment.

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