CAJ | 학술논문

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.
목적 이용핵소표기상대화절대정량(iTRAQ)기술사선유문라간균(Hp)감염상관위암적차이단백,이편심조위암적생물학표지물화약물치료파표.방법 분별수집인위암세포HGC-27급여Hp공배양24h적HGC-27적각조단백,iTRAQ시제표기후이용강리자교환주(SCX)분리,경천련질보감정급상대정량;동시통과사열Pubmed수거고중1995년1월지2014년7월공개발표적문헌,기중사용임일충차이단백질조학방법감정Hp감염상관위암적차이단백;장iTRAQ수거여문헌보도수거진행회췌분석,대획득적차이단백채용PANTHER수거고진행기인공능취류(GO)분석、PANTHER단백분류급로경(Pathway)분석.결과 통과iTRAQ표기감정출Hp감염상관위암세포HGC-27적차이단백공47개,기중하조표체적37개,상조표체적10개.통과문헌검색공납입부합요구적문헌7편,기중2개이상실험실(포괄iTRAQ실험)중복발현적단백위21개.장iTRAQ수거여문헌보도수거회췌분석득도65개차이단백,진행GO분석현시,생물학과정주요집중재세포조분중조화각충대사과정;분자공능주요집중재핵산、DNA결합,구유최화활성화전이매활성;PANTHER단백분류주요위분자반려、결합단백、골가단백、전안매화전록인자;Pathway분석현시차이단백주요삼여파금삼질병통로、조망신호통로화촉성선격소석방격소(GnRH)통로.결론 사선출다충여Hp감염상관위암적차이단백,위위암환자적조기진단제공가능적생물학표지물화약물치료적파표.
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.