武警医学2017，Vol.28Issue(7)：719-723,5.
胃癌细胞中Rab25相互作用蛋白的鉴定及功能学研究
Identification and functional analysis of Rab25-interacting proteins in gastric cancer cells
王一丞 1李园园2
作者信息
- 1. 400061,武警重庆总队医院军人病区
- 2. 116013,空军大连航空医学鉴定训练中心
- 折叠
摘要
Abstract
Objective To identify Rab25-interacting proteins in gastric cancer cells using tandem affinity purification-mass spectrometry (TAP-MS).Methods An SGC7901 cell strain that could stably express Rab25 was constructed using a StrepⅡ-6His-EGFP-Rab25 expressing plasmid by lentivirus transfection technology.After StrepⅡ-6His tandem affinity purification,mass spectrometry was carried out to analyze the interaction protein complex of Rab25.To further verify their interactions,a protein-protein network was constructed and the authenticity of interactions was verified by co-immunoprecipitation in gastric cancer cells.Results A total of 26 Rab25 interacting proteins were identified by TAP-MS technique,which were enriched in the cancer process,apoptosis,NOTCH-mediated pathway and microtubule-based movement.Co-immunoprecipitation analysis confirmed that Itgb1 interacted with Rab25,and that they collaborated to promote the invasion of SGC7901 cells according to Transwell invasion assays.Conclusions This study helps to draw a detailed map of Rab25 interactions and provides clues for treatment of gastric cancer.关键词
胃癌/Rab25/蛋白质相互作用/串联亲和纯化/质谱Key words
gastric cancer/Rab25/protein-protein interaction/tandem affinity purification/mass spectrometry分类
医药卫生引用本文复制引用
王一丞,李园园..胃癌细胞中Rab25相互作用蛋白的鉴定及功能学研究[J].武警医学,2017,28(7):719-723,5.
