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Authors: Harsh Pawar, Manoj Kumar Kashyap, Nandini A. Sahasrabuddhe, Santosh Renuse, H. C. Harsha, Praveen Kumar, Jyoti Sharma, Kumaran Kandasamy, Arivusudar Marimuthu, Bipin Nair, Sudha Rajagopalan, Jagadeesha Maharudraiah, Chennagiri Shrinivasamurthy Premalatha, Kariyanakatte Veeraiah Veerendra Kumar, M. Vijayakumar, Raghothama Chaerkady, Thotterthodi Subrahmanya Keshava Prasad, Rekha V. Kumar and Akhilesh Pandey

Harsh Pawar

Institute of Bioinformatics; International Technology Park; Bangalore, India; Rajiv Gandhi University of Health Sciences; Bangalore, India; Department of Pathology; Kidwai Memorial Institute of Oncology; Bangalore, India

Manoj Kumar Kashyap

Institute of Bioinformatics; International Technology Park; Bangalore, India

Nandini A. Sahasrabuddhe

Institute of Bioinformatics; International Technology Park; Bangalore, India; Manipal University; Manipal, India; McKusick-Nathans Institute of Genetic Medicine; Baltimore, MD USA; Department of Biological Chemistry; Johns Hopkins University School of Medicine; Baltimore, MD USA

Santosh Renuse

Institute of Bioinformatics; International Technology Park; Bangalore, India; McKusick-Nathans Institute of Genetic Medicine; Baltimore, MD USA; Department of Biological Chemistry; Johns Hopkins University School of Medicine; Baltimore, MD USA; Department of Biotechnology; Amrita Vishwa Vidyapeetham; Kollam, India

H. C. Harsha

Institute of Bioinformatics; International Technology Park; Bangalore, India

Praveen Kumar

Institute of Bioinformatics; International Technology Park; Bangalore, India

Jyoti Sharma

Institute of Bioinformatics; International Technology Park; Bangalore, India; Manipal University; Manipal, India

Kumaran Kandasamy

Institute of Bioinformatics; International Technology Park; Bangalore, India
Current address: CeMM Research Center for Molecular Medicine; Vienna, Austria

Arivusudar Marimuthu

Institute of Bioinformatics; International Technology Park; Bangalore, India; Manipal University; Manipal, India

Bipin Nair

Department of Biological Chemistry; Johns Hopkins University School of Medicine; Baltimore, MD USA

Sudha Rajagopalan

Agilent Technologies; Bangalore, India

Jagadeesha Maharudraiah

Institute of Bioinformatics; International Technology Park; Bangalore, India; RajaRajeswari Medical College; Bangalore, India

Chennagiri Shrinivasamurthy Premalatha

Department of Pathology; Kidwai Memorial Institute of Oncology; Bangalore, India

Kariyanakatte Veeraiah Veerendra Kumar

Department of Surgical Oncology; Kidwai Memorial Institute of Oncology; Bangalore, India

M. Vijayakumar

Department of Surgical Oncology; Kidwai Memorial Institute of Oncology; Bangalore, India

Raghothama Chaerkady

Institute of Bioinformatics; International Technology Park; Bangalore, India; McKusick-Nathans Institute of Genetic Medicine; Baltimore, MD USA; Department of Biological Chemistry; Johns Hopkins University School of Medicine; Baltimore, MD USA

Thotterthodi Subrahmanya Keshava Prasad

Institute of Bioinformatics; International Technology Park; Bangalore, India; Manipal University; Manipal, India; Centre of Excellence in Bioinformatics; School of Life Sciences; Pondicherry University; Pondicherry, India

Rekha V. Kumar

Department of Pathology; Kidwai Memorial Institute of Oncology; Bangalore, India

Akhilesh Pandey

Corresponding author: pandey@jhmi.edu
McKusick-Nathans Institute of Genetic Medicine; Baltimore, MD USA; Department of Biological Chemistry; Johns Hopkins University School of Medicine; Baltimore, MD USA; Department of Oncology; Johns Hopkins University School of Medicine; Baltimore, MD USA; Department of Pathology; Johns Hopkins University School of Medicine; Baltimore, MD USA

Abstract:
Esophageal squamous cell carcinoma (ESCC) is among the top ten most frequent malignancies worldwide. In this study, our objective was to identify potential biomarkers for ESCC through a quantitative proteomic approach using the isobaric tags for relative and absolute quantitation (iTRAQ) approach. We compared the protein expression profiles of ESCC tumor tissues with the corresponding adjacent normal tissue from ten patients. LC-MS/MS analysis of strong cation exchange chromatography fractions was carried out on an Accurate Mass QTOF mass spectrometer, which led to the identification of 687 proteins. In all, 257 proteins were identified as differentially expressed in ESCC as compared to normal. We found several previously known protein biomarkers to be upregulated in ESCC including thrombospondin 1 (THBS1), periostin 1 (POSTN) and heat shock 70 kDa protein 9 (HSPA9) confirming the validity of our approach. In addition, several novel proteins that had not been reported previously were identified in our screen. These novel biomarker candidates included prosaposin (PSAP), plectin 1 (PLEC1) and protein disulfide isomerase A 4 (PDIA4) that were further validated to be overexpressed by immunohistochemical labeling using tissue microarrays. The success of our study shows that this mass spectrometric strategy can be applied to cancers in general to develop a panel of candidate biomarkers, which can then be validated by other techniques.

Received: March 3, 2011; Accepted: June 7, 2011

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