Detection of epidermal growth factor receptor and human epidermal growth factor receptor 2 activating mutations in lung adenocarcinoma by high-resolution melting amplicon analysis: correlation with gene copy number, protein expression, and hormone receptor expression

Activating mutations in the epidermal growth factor receptor (EGFR) (7p12.3-p12.1) and in the human epidermal growth factor receptor 2 (HER2) (17q21.1) characterize a subset of lung carcinomas. These mutations may relate to the response of the tumor to the tyrosine kinase inhibitors gefitinib and erlotinib. High-resolution melting amplicon analysis is a screening technique that has been shown to be able to detect missense mutations as well as deletions and insertions in tumor DNA isolated from paraffin-embedded tissue sections. In this study, we used high-resolution melting amplicon analysis to screen for EGFR and human HER2 activating mutations in 39 patients with primary lung adenocarcinoma. There were 20 cases that showed bronchioloalveolar histology and 19 cases that did not. The EGFR exons screened were exons 18, 19, 20, and 21, and the HER2 exons screened were exons 19 and 20. Six (15%) of the 39 patients had tumors that contained EGFR activating mutations. Four of the mutations were in adenocarcinomas, which had some bronchioloalveolar features, and 2 mutations were in tumors without bronchioloalveolar features. The EGFR mutations were in exon 19 (2 cases), exon 20 (2 cases), and exon 21 (1 case). One case contained mutations in both exons 18 and 20. One (2.6%) of the 39 patients had a tumor that contained an HER2 activating mutation, and the mutation was located in exon 20. Two of the 6 EGFR mutation-positive cases showed polysomy for chromosome 7, and each one showed overexpression of EGFR as determined by immunohistochemical staining. The other EGFR mutation-positive cases did not show EGFR overexpression and appeared disomic for chromosome 7. The HER2 mutation-positive case was in an adenocarcinoma with bronchioloalveolar features. This tumor did not show overexpression of HER2 and was disomic for chromosome 17. For the non-EGFR mutation-positive cases, 4 (13%) of 32 evaluated cases showed polysomy for chromosome 7 and EGFR. No case showed EGFR gene amplification. Polysomy for chromosome 7 was not related to EGFR overexpression as estimated by immunohistochemistry.