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Phosphoproteome Profiling of Isogenic Cancer Cell‐Derived Exosome Reveals HSP90 as a Potential Marker for Human Cholangiocarcinoma
Author(s) -
Weeraphan Churat,
Phongdara Amornrat,
Chaiyawat Parunya,
DiskulNaAyudthaya Penchatr,
Chokchaichamnankit Daranee,
Verathamjamras Chris,
Netsirisawan Pukkavadee,
Yingchutrakul Yodying,
Roytrakul Sittiruk,
Champattanachai Voraratt,
Svasti Jisnuson,
Srisomsap Chantragan
Publication year - 2019
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201800159
Subject(s) - biology , phosphorylation , proteomics , heat shock protein , microvesicles , exosome , protein phosphorylation , blot , hsp27 , cancer research , biomarker , proteome , cancer cell , cancer , microbiology and biotechnology , hsp70 , bioinformatics , biochemistry , microrna , genetics , gene , protein kinase a
The northeastern region of Thailand is well known to have a high incidence and mortality of cholangiocarcinoma (CCA). Protein phosphorylation status has been reported to reflect a key determinant of cellular physiology, but identification of phosphoproteins can be a problem due to the presence of phosphatase. Exosomes are stable toward circulating proteases and other enzymes in human blood and can be recognized before the onset of cancer progression. Here an in vitro metastatic model of isogenic CCA cells is used to provide insight into the phosphorylation levels of exosomal proteins derived from highly invasive cells. Gel‐based and gel‐free proteomics approaches are used to reveal the proteins differentially phosphorylated in relation to tumor cell phenotypes. Forty‐three phosphoproteins are identified with a significant change in phosphorylation level. Phos‐tag western blotting and immunohistochemistry staining are then employed to validate the candidate phosphoproteins. Heat shock protein 90 is successfully confirmed as being differentially phosphorylated in relation to tumor malignancy. Importantly, the aberrant phosphorylation of exosomal proteins might serve as a promising tool for the development of a biomarker for metastatic CCA.