Identification of Candidate Biomarkers for Early Detection of Human Lung Squamous Cell Cancer by Quantitative Proteomics

• Type of Material: Article
• Authors: Gu-Qing Zeng, Central South University; Pang-Fei Zhang, Central South University; Xingming Deng, Emory University; Feng-Lei Yu, Central South University; Cui Li, Central South University; Yan Xu, Central South University; Hong Yi, Central South University; Mao-Yu Li, Central South University; Rong Hu, Central South University; Jian-Hong Zuo, Central South University; Xin-Hui Li, Central South University; Xun-Xun Wan, Central South University; Jia-Quan Qu, Central South University; Qiu-Yan He, Central South University; Jian-Huang Li, Central South University; Xu Ye, Central South University; Yu Chen, Central South University; Jiao-Yang Li, Central South University; Zhi-Qiang Xiao, Central South University
• Language: English
• Date: 2012-06-01
• Publisher: American Society for Biochemistry and Molecular Biology
• Publication Version: Final Published Version
• Copyright Statement: © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.
• Final Published Version (URL): http://dx.doi.org/10.1074/mcp.M111.013946
• Title of Journal or Parent Work: Molecular & Cellular Proteomics
• Volume: 11
• Issue: 6
• Start Page: M111.013946
• End Page: M111.013946
• Grant/Funding Information: This work was supported by National Nature Science Foundation of China (30973290, 81172559, 81172302), Lotus Scholars Program of Hunan Province, China (2007–362), Key Research Program from Science and Technology Department of Hunan Province, China (2010FJ2009), Nature Science Foundation of Hunan Province, China (11JJ2045), Scientific Research Fund of Hunan Provincial Education Department, China (09C837), and Scientific Research Fund of Hunan Provincial Health Department, China (B2010-037).
• Keywords: Life Sciences & Biomedicine; Multidimensional Liquid-chromatography; Science & Technology; Nasopharyngeal Carcinoma; Laser capture microdissection; HSP27; Biochemical Research Methods; Tandem mass-spectronomy; Expression; Heat shock proteins; Serum; Creatine; Glutathione S-transferase; Biochemistry & Molecular Biology
• Research Categories: Biology, Cell; Chemistry, Biochemistry; Biology, Molecular; Health Sciences, Oncology

Abstract

To discover novel biomarkers for early detection of human lung squamous cell cancer (LSCC) and explore possible mechanisms of LSCC carcinogenesis, iTRAQ-tagging combined with two dimensional liquid chromatography tandem MS anal ysis was used to identify differentially expressed proteins in human bronchial epithelial carcinogenic process using laser capture microdissection-purified normal bronchial epithelium (NBE), squamous metaplasia (SM), atypical hyperplasia (AH), carcinoma in situ (CIS) and invasive LSCC. As a result, 102 differentially expressed proteins were identified, and three differential proteins (GSTP1, HSPB1 and CKB) showing progressively expressional changes in the carcinogenic process were selectively validated by Western blotting. Immunohistochemistry was performed to detect the expression of the three proteins in an independent set of paraffin-embedded archival specimens including various stage tissues of bronchial epithelial carcinogenesis, and their ability for early detection of LSCC was evaluated by receiver operating characteristic analysis. The results showed that the combination of the three proteins could perfectly discriminate NBE from preneoplastic lesions (SM, AH and CIS) from invasive LSCC, achieving a sensitivity of 96% and a specificity of 92% in discriminating NBE from preneoplatic lesions, a sensitivity of 100% and a specificity of 98% in discriminating NBE from invasive LSCC, and a sensitivity of 92% and a specificity of 91% in discriminating preneoplastic lesions from invasive LSCC, respectively. Furthermore, we knocked down GSTP1 in immortalized human bronchial epithelial cell line 16HBE cells, and then measured their susceptibility to carcinogen benzo(a)pyrene-induced cell transformation. The results showed that GSTP1 knockdown significantly increased the efficiency of benzo(a)pyrene-induced 16HBE cell transformation. The present data first time show that GSTP1, HSPB1 and CKB are novel potential biomarkers for early detection of LSCC, and GSTP1 down-regulation is involved in human bronchial epithelial carcinogenesis.