4-氨基-2-三氟甲基苯基维甲酸酯对人白血病K562细胞的蛋白质组学研究

The proteomics research of 4-amino-2-trifluoromethyl-phenyl retinate on human leukemia K562 cells

目的研究新型维甲酸衍生物4-氨基-2-三氟甲基苯基维甲酸酯(4-amino-2-trifluoromethyl-phenyl retinate,ATPR)诱导人白血病K562细胞分化作用的蛋白质组学机制。方法1×10-6mol·L-1的ATPR及ATRA分别作用于人白血病K562细胞48 h后,收集细胞并提取总蛋白,纯化之后使用胰蛋白酶酶解,固相萃取法脱盐,高分辨率液相色谱质谱联用仪检测肽段,使用Proteome Discoverer 1.2软件寻找出差异表达的蛋白质,利用生物信息学DAVID数据库、KEGG数据库、STRING数据库,分析鉴定出的差异蛋白质所具有的分子功能、所参与的生物学过程等信息。结果 ATPR组特异性的蛋白质鉴定出120个,ATRA组特异性蛋白质有143个,两者共有蛋白422个。DAVID分析显示ATPR特异性蛋白质主要参与39个生物学过程,包括蛋白质和大分子的代谢、蛋白质转运和定位等。KEGG分析发现,ATPR组特异性蛋白质主要参与新陈代谢过程、PI3K-Akt信号通路、TGF-β信号通路等其他癌症相关信号通路。STRING蛋白相互作用网络分析显示ATPR特异性蛋白质,如EIF3A、EIF6、RPL3、RPL8、RPL13、RPL7A、RPL21、RPS3、RPS14、NACA、BTF3、NHP2L1、PPP2CA蛋白与其他≥10个相关蛋白存在直接相互作用关系。结论 ATPR组特异性中心蛋白质均参与对细胞生长增殖、诱导细胞分化和凋亡等过程的调控,这些蛋白质间的相互作用网络及特异性中心蛋白质是ATPR诱导K562细胞分化作用的可能机制。

Aim To explore the proteomics mechanism of the differentiation induction effect of 4-amino-2-trifluoromethyl-phenyl retinate( ATPR) on human leukemia K562 cells. Methods Human leukemia K562 cells were incubated with the same concentration( 1 × 10- 6mol·L- 1) of ATPR or ATRA for 48 hours. The total cell proteins were collected,purified and digested by trypsin,solid phase extraction,and the peptides were detected by ESI-LC-MS / MS. The difference of the protein expression between the cells treated with ATPR and ATRA was compared by using the Discoverer Proteome 1. 2 software,and the molecular function,the biological process and other information of those proteins were analyzed based on the DAVID, KEGG,STRING databases. Results 120 specific proteins were identified only in the ATPR group,143 only in the ATRA group, and 422 other proteins in both groups. Results of DAVID analysis showed that ATPRinduced specific proteins were mainly involved in 39 biological processes of proteins and macromolecules metabolism,protein transport and localization and so on. Results of KEGG analysis revealed that ATPR-induced proteins participated in signal pathways,mainly metabolic pathways,PI3K-Akt signal pathway,TGFbeta signal pathway and other pathways in cancer.String protein interaction network analysis displayed that ATPR-induced proteins, like EIF3 A, EIF6,RPL3, RPL8, RPL13, RPL7 A, RPL21, RPS3,RPS14,NACA,BTF3,NHP2L1,PPP2 CA proteins had direct interactions with more than or equal to 10 associated proteins. Conclusion The differentiation induction effect of ATPR on K562 cells might be ascribed to the ATPR-induced proteins interaction network and the specific central proteins it induced,which are involved in the regulation of cell proliferation,differentiation and apoptosis.