采用IP-XL/MS技术研究SGC7901胃癌细胞中EGFR信号通路动态变化  

Dynamic changes in EGFR signaling pathway in SGC7901 gastric cancer cells by IP-XL-MS technology

作　　者：汲淑慧 王旖旎 汪宜 秦钧 JI Shu-hui;WANG Yi-ni;WANG Yi;QIN Jun(State Key Laboratory of Proteomics,Beijing Proteome Research Center,National Center for Protein Sciences Beijing,Institute of Lifeomics,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 102206,China)

机构地区：[1]军事科学院军事医学研究院生命组学研究所国家蛋白质科学中心(北京)北京蛋白质组研究中心蛋白质组学国家重点实验室,北京102206

出　　处：《军事医学》2018年第4期241-248,共8页Military Medical Sciences

基　　金：国家自然科学基金资助项目(31770892)

摘　　要：目的研究胃癌细胞在表皮生长因子(EGF)刺激下表皮生长因子受体(EGFR)相互作用蛋白的时序性变化。方法筛选高表达EGFR的胃癌细胞系;EGF刺激不同时间后,利用甲醛固定免疫沉淀联合质谱(IP-XL/MS)技术对胃癌细胞中EGFR相互作用蛋白表达谱进行无标定量,捕获EGFR网络的动态变化。结果与结论 8种胃癌细胞系中,SGC7901细胞表达EGFR量最高。4次生物学重复实验,共鉴定到3728个蛋白,625个EGFR相互作用蛋白,包括已知的相互作用蛋白(GRB2、CBL、SHC1等),并发现59个新蛋白(ANKFY、LGR4、SNX3、VPS26A、ZFYVE20等)。ANOVA检验分析得到406个响应EGF刺激的差异表达蛋白,根据动力学变化可分成5个簇,其具有不同的生物学功能,如蛋白转运、內吞、囊泡循环和蛋白酶体输运等,共同参与EGFR信号传递的动态调控,交叉调节多条信号通路。Objective To study the temporal changes in epidermal growth factor receptor（EGFR） and associated proteins in gastric cancer cells stimulated by EGF. Methods We began by sorting the cell line which overexpressed the EGFR in gastric cancer cell lines available at our laboratory. After that,the dynamics of the EGFR network was captured by quantifying the expression profile of EGFR-interacting proteins using IP-XL-MS technique in SGC7901 cells stimulated with EGF at different time gradients. Results and Conclusion SGC7901 cells expressed the highest amount of EGFR. A total of 3728 proteins were identified by mass spectrometry at four biological replicates on EGF stimulation,and 625 EGFR interacting proteins were obtained,including many known interactions（GRB2,CBL,SHC1,et al） and 59 new proteins（ANKFY1,CLTB,LGR4,SNX3,VPS26 A,ZFYVE20,et al）. Data analyses based on ANOVA analysis discovered 406 proteins that were differentially expressed in response to EGF exposure,which were classified into 5 clusters according to their different dynamics. These proteins have different protein functions,such as protein transport,endocytosis,vesicle cycle and proteasome transport,but they are also involved in the dynamic regulation of EGFR signaling and cross regulation of multiple signal paths.

关 键 词：胃肿瘤 表皮生长因子 免疫沉淀法 IP-XL/MS 蛋白质转运 动态变化 

分 类 号：R735.2[医药卫生—肿瘤]