Overexpression of EGFR in head and neck squamous cell carcinoma is associated with inactivation of SH3GL2 and CDC25A genes

PLoS One. 2013 May 10;8(5):e63440. doi: 10.1371/journal.pone.0063440. Print 2013.

Abstract

The aim of this study is to understand the mechanism of EGFR overexpression in head and neck squamous cell carcinoma (HNSCC). For this reason, expression/mutation of EGFR were analyzed in 30 dysplastic head and neck lesions and 148 HNSCC samples of Indian patients along with 3 HNSCC cell lines. In addition, deletion/methylation/mutation/expression of SH3GL2 (associated with EGFR degradation) and CDC25A (associated with dephosphorylation of EGFR) were analyzed in the same set of samples. Our study revealed high frequency of EGFR overexpression (66-84%), low frequency of gene amplification (10-32.5%) and absence of functional mutation in the dysplastic lesions and HNSCC samples. No correlation was found between protein overexpression and mRNA expression/gene amplification status of EGFR. On the other hand, frequent alterations (deletion/methylation) of SH3GL2 (63-77%) and CDC25A (37-64%) were seen in the dysplastic and HNSCC samples. Two novel single nucleotide polymorphism (SNPs) were found in the promoter region of SH3GL2. Reduced expression of these genes showed concordance with their alterations. Overexpression of EGFR and p-EGFR were significantly associated with reduced expression and alterations of SH3GL2 and CDC25A respectively. In-vitro demethylation experiment by 5-aza-2'-deoxycytidine (5-aza-dC) showed upregulation of SH3GL2 and CDC25A and downregulation of EGFR expression in Hep2 cell line. Poor patient outcome was predicted in the cases with alterations of SH3GL2 and CDC25A in presence of human papilloma virus (HPV) infection. Also, low SH3GL2 and high EGFR expression was a predictor of poor patient survival. Thus, our data suggests that overexpression of EGFR due to its reduced degradation and dephosphorylation is needed for development of HNSCC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics*
  • Adaptor Proteins, Signal Transducing / metabolism
  • Adult
  • Aged
  • Carcinoma, Squamous Cell / genetics*
  • Carcinoma, Squamous Cell / mortality
  • Carcinoma, Squamous Cell / pathology
  • Carcinoma, Squamous Cell / virology
  • DNA Methylation
  • ErbB Receptors / genetics*
  • ErbB Receptors / metabolism
  • Gene Amplification
  • Gene Deletion
  • Gene Expression Regulation, Neoplastic*
  • Gene Expression*
  • Gene Silencing
  • Head and Neck Neoplasms / genetics*
  • Head and Neck Neoplasms / mortality
  • Head and Neck Neoplasms / pathology
  • Head and Neck Neoplasms / virology
  • Humans
  • Middle Aged
  • Mutation
  • Papillomaviridae
  • Promoter Regions, Genetic
  • Reproducibility of Results
  • Squamous Cell Carcinoma of Head and Neck
  • cdc25 Phosphatases / genetics*
  • cdc25 Phosphatases / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • SH3GL2 protein, human
  • ErbB Receptors
  • CDC25A protein, human
  • cdc25 Phosphatases

Grants and funding

Financial support for this work was provided by grants from Department of Biotechnology, (BT/PR/5524/Med/14/649/2004), Government of India, to Dr. CK Panda and Dr. S Roychoudhury, Council for Scientific and Industrial Research-Project (IAP-001), Government of India to Dr. S Roychoudhury and CSIR-SRF Fellowship (grant no. 09/30 (0053)2k9-EMR-I) to Mr. GP Maiti. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.