Poster Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

Phosphoproteome profiling of isogenic cancer cell-derived exosome reveals HSP90 as a potential marker for human cholangiocarcinoma (#909)

Churat Weeraphan 1 , Amornrat Pongdara 2 3 , Parunya Chaiyawat 4 , Penchatr Diskul-Na-Ayudthaya 5 , Daranee Chokchaichamnankit 6 , Chris Verathamjamras 6 , Pukkavadee Netsirisawan 6 , Yodying Yingchutrakul 7 , Sittiruk Roytrakul 7 , Voraratt Champattanachai 6 , Jisnuson Svasti 4 6 , Chantragan Srisomsap 6
  1. Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Songkhla, Thailand
  2. Faculty of Medical Technology, Prince of Songkla University, Songkhla , Thailand
  3. Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University, Songkhla , Thailand
  4. Applied Biological Sciences Program, Chulabhorn Graduate Institute, Bangkok, Thailand
  5. Soonchunhyang Institute of Medi-bio Science (SIMS) , Soonchunhyang University, Chungcheongnam-do, Republic of Korea
  6. Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok , Thailand
  7. Proteomics Research Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, Thailand

The Northeastern region of Thailand is well known to have a high incidence and mortality of cholangiocarcinoma. 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. Growing evidence indicates that exosomes are stable towards circulating proteases and other enzymes in human blood and can be recognized before the onset of cancer progression.Here an in vitrometastatic model of isogenic cholangiocarcinoma cells was used to provide insight into the phosphorylation levels of exosomal proteins derived from highly invasive cells. Gel-based and gel-free proteomics approaches were used to reveal the proteins differentially phosphorylated in relation to tumor cell phenotypes. Forty-three phosphoproteins were identified with a significant change in phosphorylation level. Phos-tag western blotting and Immunohistochemistry staining was then employed to validate the candidate phosphoproteins. Heat shock protein 90 was 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 cholangiocarcinoma.

  1. Weeraphan C, Phongdara A, Chaiyawat P, Diskul-Na-Ayudthaya P, Chokchaichamnankit D, Verathamjamras C, et al. Phosphoproteome Profiling of Isogenic Cancer Cell-Derived Exosome Reveals HSP90 as a Potential Marker for Human Cholangiocarcinoma. Proteomics. 2019
  2. Chen IH, Xue L, Hsu CC, Paez JS, Pan L, Andaluz H, et al. Phosphoproteins in extracellular vesicles as candidate markers for breast cancer. Proc Natl Acad Sci U S A. 2017;114(12):3175-80.