Elsevier

Oral Oncology

Volume 51, Issue 1, January 2015, Pages 53-58
Oral Oncology

Rare occurrence of EGFRvIII deletion in head and neck squamous cell carcinoma

https://doi.org/10.1016/j.oraloncology.2014.08.014Get rights and content

Highlights

  • Out of 638 samples analyzed, only 2 samples were positive for EGFRvIII deletion.

  • EGFRvIII deletion is very rare in head and neck squamous cell carcinoma (HNSCC).

  • EGFRvIII deletion should not be included in regular diagnostic tests for HNSCC.

Summary

Background

The epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor and is overexpressed in up to 90% of head and neck squamous cell carcinoma (HNSCC) cases. The EGFR truncation mutation, EGFR variant III (EGFRvIII), harbors an in-frame deletion of exons 2–7 (801 bp) that leads to the constitutive activation of downstream signaling. EGFRvIII has been reported in ∼40% of glioblastomas (GBM), but its presence in HNSCC remains controversial.

Methods

EGFRvIII deletion in 638 HNSCC samples was analyzed using: (i) quantitative Real-Time polymerase chain reaction (qRT-PCR) on 108 HNSCC samples with direct detection of the EGFRvIII breakpoint, (ii) RNA-Seq analysis on 7 HNSCC tumor tissues and 425 The Cancer Genome Atlas (TCGA) HNSCC samples, and (iii) immunohistochemistry (IHC) for EGFRvIII using an established antibody (L8A4) on a tissue microarray of 105 HNSCC samples.

Results

qRT-PCR did not show the presence of EGFRvIII in any of the samples analyzed. Furthermore, we could not detect any EGFRvIII transcripts in the RNA-Seq data of the seven HNSCC samples. However, 2 samples out of 425 TCGA HNSCC samples had EGFRvIII specific reads. EGFRvIII IHC results were assessed as negative for all samples.

Conclusion

Our results firmly establish that EGFRvIII is very rare in HNSCC as only 2 out of 638 (0.31%) samples we analyzed overall, or 2 out of 540 (0.37%) using mRNA based approaches, were positive for EGFRvIII. EGFRvIII is extremely rare in HNSCC and the clinical significance remains unclear. We propose not to include EGFRvIII testing in regular diagnostic tests for HNSCC.

Section snippets

Background

The epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor and one of the most commonly affected growth factor receptors in cancer [1], [2]. EGFR is overexpressed in several epithelial cancers, including squamous cell carcinoma of the head and neck (HNSCC) in which it exhibits overexpression in up to 90% of tumors [2], [3], [4], [5]. Elevated levels of EGFR are usually induced by amplification of the EGFR gene or through altered transcriptional regulation and

Nucleic acid extraction from HNSCC samples

A total of 108 advanced HNSCC specimens were selected for the qRT-PCR analysis. Of these 108 samples, 56 were formalin fixed and paraffin embedded (FFPE) and 52 were Optimal Cutting Temperature compound (OCT) frozen. Samples were obtained from the Head and Neck Cancer tissue bank of the University of Chicago (IRB approved protocol UCCCC#8980). The clinical details of OCT frozen samples is given in Table 1. The FFPE samples were de-identified and their details are not available to us. A section

Results

In the present study we analyzed the presence of EGFRvIII in 638 HNSCC samples using rigorous gold-standard methodologies. qRT-PCR was used for 108 HNSCC samples. Around half of the samples were fresh frozen (52 samples) whereas the other half was FFPE (56 samples). In all these samples including the positive control we could successfully amplify the wild-type EGFR as well as GAPDH. However, none of the samples showed amplification of EGFRvIII when they were analyzed following methodology of

Discussion

The epidermal growth factor receptor (EGFR) is a growth-promoting molecule and it has long been associated with increased tumorogenesis [1]. EGFRvIII was first detected in glioblastoma [23] and was later reported to exist in other cancer types of epithelial origin [6], [7], [8], [9]; however, its presence in HNSCC remains controversial.

EGFRvIII in HNSCC was first described by Sok et al. (2006) [10], who found it in 42% of HNSCC tumors. EGFRvIII always appeared in conjunction with the wild-type

Conclusion

Our results establish that EGFRvIII expression is exceedingly rare in HNSCC as only 2 samples out of 638 (0.31%) samples or 2 samples out of 540 (0.37%) samples using mRNA based approaches were positive. We propose not to include EGFRvIII in regular diagnostic tests for the HNSCC given its’ rarity, and indeterminate biologic meaning for HNSCC.

Conflict of interest statement

None declared.

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