4-氨基-2-三氟甲基苯基维甲酸酯对人胃癌SGC-7901细胞的磷酸化蛋白质组学研究

摘要

Objective To investigate levels of phosphorylation of protein when the new retinoid derivatives 4-Ami-no-2-Trifluoromethyl-Phenyl Retinate (ATPR)on gastric cancer SGC-7901 cells,which could help to understand the possible mechanism of differentiation,and found the targets by phosphoproteomics.Methods Gastric cancer SGC-7901 cells were incubated for 48 hours with ATPR,collected and extracted of total cell protein,digested by trypsin,phosphopeptides were enriched by TiO2 beads and analyzed by LC-MS /MS.Used Proteome Discoverer1.2 to find different proteins and phosphosites.Bioinformatic was applied to analyze the function and subcellular locali-zation of phosphorylated proteins to explain these different proteins by DAVID,KEGG,IPA software.Results We identified 109 proteins,including 45 phosphoproteins and 121 phosphosites by analysis of MS.15 proteins ap-peared on the ATPR group and 20 proteins in the control group.DAVID analysis results displayed that different phosphorproteins were involved in regulating cellular process,biological process,gene expression,metabolic process and so on;the molecular functions of different phosphoproteins may include protein binding and nucleic acid binding.ERBB signaling pathway,RNA transport,Protein processing in endoplasmic reticulum,p53 signa-ling pathway were involved by KEGG analysis.IPA software results displayed that the ubiquitin ligase (UBC)be-came associated protein of several proteins.Conclusion ATPR can induce differentiation of gastric cancer cell SGC-7901,the mechanism may be the results of interaction of proteins or genes,such as HDGF,CAST,ABCG4 and CAP1,and regulate the phosphorylation of these proteins.%目的：研究观察新型维甲酸衍生物4-氨基-2-三氟甲基苯基维甲酸酯（ATPR）作用于胃癌 SGC-7901细胞后蛋白质的磷酸化情况来了解 ATPR 诱导分化作用的可能机制和作用靶点。方法 ATPR 作用于胃癌 SGC7901细胞48 h 后，收集并提取总蛋白，通过胰蛋白酶酶解，TiO2磁珠法富集磷酸肽，高分辨率液质联用仪器检测磷酸肽，使用 Proteome Discoverer1．2软件寻找差异的蛋白质和磷酸化位点，利用生物信息学网站和 DAVID、KEGG、IPA 软件，分析这些磷酸化蛋白质的功能及亚细胞定位等信息。结果最终鉴定出109个蛋白质，其中45个磷酸化蛋白质，共有121个磷酸化位点，差异蛋白中有15个仅出现在 ATPR 组，20个仅出现在正常组。DAVID 分析结果显示差异磷酸化的蛋白质参与调控细胞进程、生物进程、基因表达、细胞的代谢过程等；KEGG分析结果显示差异磷酸化蛋白质主要有参与 ERBB 信号通路、RNA 的转运、内质网蛋白加工过程、p53信号通路等。IPA 软件分析一些差异蛋白质之间的关联蛋白是泛素连接酶（UBC）。结论 ATPR 对胃癌细胞 SGC-7901细胞诱导分化的机制可能是通过作用于多种蛋白质或基因，如肝癌衍生生长因子（HDGF）、钙蛋白酶抑制蛋白（CAST）、ATP 结合盒G 亚族蛋白4（ABCG4）和腺苷酸环化酶关联蛋白1（CAP1）而发挥作用，并通过影响这些蛋白质的磷酸化从而实现 AT-PR 对胃癌细胞的诱导分化作用。