Identification and functional analysis of Rab25-interacting proteins in gastric cancer cells
WANG Yicheng1 and LI Yuanyuan2
1. Department of Servicemen,Chongqing Municipal Corps Hospital,Chinese People’s Armed Police Force, Chongqing 400061,China; 2.Air Force Aviation Medicine Identification and Training Center, Dalian 116013, China
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.
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.
WANG Yicheng and LI Yuanyuan. Identification and functional analysis of Rab25-interacting proteins in gastric cancer cells. Med. J. Chin. Peop. Armed Poli. Forc., 2017, 28(7): 719-723.
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