SILAC-based quantitative proteomic analysis of gastric cancer secretome

Arivusudar Marimuthu, Yashwanth Subbannayya, Nandini A. Sahasrabuddhe, Lavanya Balakrishnan, Nazia Syed, Nirujogi Raja Sekhar, Teesta V. Katte, Sneha M. Pinto, Srinivas M. Srikanth, Praveen Kumar, Harsh Pawar, Manoj K. Kashyap, Jagadeesha Maharudraiah, Hassan Ashktorab, Duane T. Smoot, Girija Ramaswamy, Rekha V. Kumar, Yulan Cheng, Stephen J. Meltzer, Juan Carlos RoaRaghothama Chaerkady, T. S.Keshava Prasad, H. C. Harsha, Aditi Chatterjee, Akhilesh Pandey

Research output: Contribution to journalArticle

Abstract

Purpose: Gastric cancer is a commonly occurring cancer in Asia and one of the leading causes of cancer deaths. However, there is no reliable blood-based screening test for this cancer. Identifying proteins secreted from tumor cells could lead to the discovery of clinically useful biomarkers for early detection of gastric cancer. Experimental design: A SILAC-based quantitative proteomic approach was employed to identify secreted proteins that were differentially expressed between neoplastic and non-neoplastic gastric epithelial cells. Proteins from the secretome were subjected to SDS-PAGE and SCX-based fractionation, followed by mass spectrometric analysis on an LTQ-Orbitrap Velos mass spectrometer. Immunohistochemical labeling was employed to validate a subset of candidates using tissue microarrays. Results: We identified 2205 proteins in the gastric cancer secretome of which 263 proteins were overexpressed greater than fourfold in gastric cancer-derived cell lines as compared to non-neoplastic gastric epithelial cells. Three candidate proteins, proprotein convertase subtilisin/kexin type 9 (PCSK9), lectin mannose binding 2 (LMAN2), and PDGFA-associated protein 1 (PDAP1) were validated by immunohistochemical labeling. Conclusions and clinical relevance: We report here the largest cancer secretome described to date. The novel biomarkers identified in the current study are excellent candidates for further testing as early detection biomarkers for gastric adenocarcinoma.

Original languageEnglish (US)
Pages (from-to)355-366
Number of pages12
JournalProteomics - Clinical Applications
Volume7
Issue number5-6
DOIs
StatePublished - Jun 2013

Keywords

  • Biomarkers
  • Cell supernatants
  • Early diagnosis
  • Gastric carcinoma
  • In vivo labeling
  • Secreted proteins

ASJC Scopus subject areas

  • Clinical Biochemistry

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  • Cite this

    Marimuthu, A., Subbannayya, Y., Sahasrabuddhe, N. A., Balakrishnan, L., Syed, N., Sekhar, N. R., Katte, T. V., Pinto, S. M., Srikanth, S. M., Kumar, P., Pawar, H., Kashyap, M. K., Maharudraiah, J., Ashktorab, H., Smoot, D. T., Ramaswamy, G., Kumar, R. V., Cheng, Y., Meltzer, S. J., ... Pandey, A. (2013). SILAC-based quantitative proteomic analysis of gastric cancer secretome. Proteomics - Clinical Applications, 7(5-6), 355-366. https://doi.org/10.1002/prca.201200069