Phosphoproteomics to analyze PTPLAD1-regulated tyrosine-phosphorylated proteins in colon cancer cells
HU Yang, YANG Jie, YU Ru-yuan, WANG Yang
Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, College of Life Science and Technology, Jinan University, Guangzhou 510632, China
AIM: To identify and analyze tyrosine-phosphorylated proteins regulated by protein tyrosine phosphatase-like A domain containing protein 1 (PTPLAD1) in colon cancer cells by phosphoproteomics. METHODS: The expression of PTPLAD1 in colon cancer cell line HCT-116 was knocked down by small interfering RNAs, and the differential expression of tyrosine-phosphorylated proteins in response to the konckdown of PTPLAD1 in HCT-116 cells was identified by stable isotope labeling with amino acid in cell culture (SILAC), coupled with the tyrosine phosphorylation antibody immunoprecipitation and LC-MS/MS analysis. The Ingenuity Pathway Analysis (IPA) software was employed for bioinformatics analysis on the differentially-expressed proteins. RESULTS: A total of 20 differentially-expressed tyrosine-phosphorylated proteins were identified by MS, including 8 markedly up-regulated and 10 evidently down-regulated proteins. IPA software suggested that these proteins were mainly associated with the disease of cancer, tissue development and function, and cell death and survival. CONCLUSION: We successfully identified PTPLAD1-regulated differentially-expressed tyrosine-phosphorylated proteins in colon cancer cell line HCT-116. Our analysis suggests that PTPLAD1-regulated proteins in colon cancer are closely correlated with colon cancer.
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