Keywords

Gastric Cancer, Rkip, Affinity Purification, Proteomics, 14-3-3 Protein

Abstract

Abnormal expression of raf kinase inhibitory protein (rkip) plays an important role in the occurrence and development of gastric cancer. In order to elucidate its mechanism of action, liposomes were used to transfect rkip-3xflag-pcdna3.1 plasmid into sgc7901 cells to establish rkip-3xflag Highly expressed sgc7901 cells; and used 3xflag tag affinity chromatography technology combined with mass spectrometry analysis to isolate and identify proteins that interact with rkip, and applied co-immunoprecipitation combined with western-blot to further verify the mass spectrometry results. A total of 66 rkip-interacting proteins were identified, and their functional categories include protein metabolism enzymes, biological oxidation-related enzymes, cytoskeletal proteins, molecular chaperones, signal transduction-related proteins, enzymatic hydrolysis-related proteins, etc. And it was confirmed for the first time that the 14-3-3 protein interacts with rkip. It provides important clues to elucidate the mechanism of rkip in the occurrence and development of gastric cancer, and provides new targets for early diagnosis, treatment and prognosis monitoring of gastric cancer.

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Original research done by Gu Huan, Yan Lu, Li Jia, Zhang Guiying

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