SILAC-based quantitative proteomic approach to identify potential biomarkers from the esophageal squamous cell carcinoma secretome
Year: 2010
Type: article
Abstract: The identification of secreted proteins that are differentially expressed between non-neoplastic and esophageal squamous cell carcinoma (ESCC) cells can provide potential biomarkers of ESCC. We used a SILAC-based quantitative proteomic approach to compare the secretome of ESCC cells with that of non-neoplastic esophageal squamous epithelial cells. Proteins were resolved by SDS-PAGE, and tandem mass spectrometry analysis (LC-MS/MS) of in-gel trypsin-digested peptides was carried out on a high-accuracy qTOF mass spectrometer. In total, we identified 441 proteins in the combined secretomes, including 120 proteins with > 2-fold upregulation in the ESCC secretome vs. that of non-neoplastic esophageal squamous epithelial cells. In this study, several potential protein biomarkers previously known to be increased in ESCC including matrix metalloproteinase 1, transferrin receptor, and transforming growth factor beta-induced 68 kDa were identified as overexpressed in the ESCC-derived secretome. In addition, we identified several novel proteins that have not been previously reported to be associated with ESCC. Among the novel candidate proteins identified, protein disulfide isomerase family a member 3 (PDIA3), GDP dissociation inhibitor 2 (GDI2), and lectin galactoside binding soluble 3 binding protein (LGALS3BP) were further validated by immunoblot analysis and immunohistochemical labeling using tissue microarrays. This tissue microarray analysis showed overexpression of protein disulfide isomerase family a member 3, GDP dissociation inhibitor 2, and lectin galactoside binding soluble 3 binding protein in 93%, 93% and 87% of 137 ESCC cases, respectively. Hence, we conclude that these potential biomarkers are excellent candidates for further evaluation to test their role and efficacy in the early detection of ESCC.
Source: Cancer Biology & Therapy
Authors: Manoj Kumar Kashyap, H. C. Harsha, Santosh Renuse, Harsh Pawar, Nandini A. Sahasrabuddhe, Min-Sik Kim, Arivusudar Marimuthu, Shivakumar Keerthikumar, Babylakshmi Muthusamy, Kumaran Kandasamy, Yashwanth Subbannayya, Thottethodi Subrahmanya Keshava Prasad, Riaz Mahmood, Raghothama Chaerkady, Stephen J. Meltzer, Rekha V. Kumar, Anil K. Rustgi, Akhilesh Pandey
Institutions: Johns Hopkins University, Kuvempu University, Johns Hopkins Medicine, Institute of Bioinformatics, Amrita Vishwa Vidyapeetham, Rajiv Gandhi University of Health Sciences, Manipal Academy of Higher Education, Kidwai Memorial Institute of Oncology, Abramson Cancer Center, University of Pennsylvania
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: bronze
Funders: Department of Biotechnology, Ministry of Science and Technology, India, Indian Council of Medical Research, Council of Scientific and Industrial Research, India, University Grants Commission, Medical Research Council, National Cancer Institute, National Institute of Diabetes and Digestive and Kidney Diseases
OpenAlex ID: W1969407277