Phosphoproteomics to analyze PTPLAD1-regulated tyrosine-phosphoryla-ted proteins in colon cancer cells
10.3969/j.issn.1000-4718.2015.05.014
- VernacularTitle:磷酸化蛋白质组学鉴定 PTPLAD1调控的结肠癌细胞酪氨酸磷酸化蛋白质
- Author:
Yang HU
;
Jie YANG
;
Ruyuan YU
;
Yang WANG
- Publication Type:Journal Article
- Keywords:
Protein tyrosine phosphatase-like A domain containing protein 1;
HCT-116 cells;
Small inter-fering RNA;
Stable isotope labeling with amino acid;
Phosphoproteomics
- From:
Chinese Journal of Pathophysiology
2015;(5):845-851
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To identify and analyze tyrosine-phosphorylated proteins regulated by protein tyrosine phos-phatase-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 differenti-al 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 im-munoprecipitation and LC-MS/MS analysis.The Ingenuity Pathway Analysis ( IPA) software was employed for bioinformat-ics analysis on the differentially-expressed proteins.RESULTS:A total of 20 differentially-expressed tyrosine-phosphoryla-ted proteins were identified by MS, including 8 markedly up-regulated and 10 evidently down-regulated proteins.IPA soft-ware 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.
