J Breast Cancer.  2017 Jun;20(2):150-159. 10.4048/jbc.2017.20.2.150.

Mutations of the Epidermal Growth Factor Receptor Gene in Triple-Negative Breast Cancer

• Kim A
• Jang MH
• Lee SJ
• Bae YK

• KMID: 2389752
• DOI: http://doi.org/10.4048/jbc.2017.20.2.150

Abstract

PURPOSE
Epidermal growth factor receptor (EGFR) is considered a potential therapeutic target for anti-EGFR therapy in triple-negative breast cancer (TNBC). However, the frequency of EGFR gene mutation in TNBC is low and varies with ethnicity. This study aimed to investigate the incidence of EGFR gene mutation in TNBC.
METHODS
EGFR protein expression was evaluated by immunohistochemistry in tissue microarrays of 493 TNBC cases using four different primary antibodies, which included mutation-specific antibodies. For cases with an immunoreactivity level ≥1+, we performed pyrosequencing analysis for EGFR gene mutation. A case was considered mutation-positive when its mutation frequency minus its limit of detection (LOD) was >10%. Cases with mutation frequency higher than LOD were assessed for EGFR gene mutation status using the Cobas assay and by peptide nucleic acid-mediated polymerase chain reaction (PNA-clamping).
RESULTS
Among 493 TNBCs, 148 (30.0%) exhibited staining ≥1+ for EGFR, including 78 with 1+, 49 with 2+, and 21 with 3+. Positive EGFR expression (≥2+) was significantly associated with lymphovascular invasion (p=0.010), but not with overall survival (p=0.444) or disease-free survival (p=0.388). None of the 493 TNBCs harbored an EGFR gene mutation. Among 148 cases with an EGFR staining result ≥1+, five (3.4%) showed mutation frequencies (4.4%-10.9%) higher than LOD (2.6%-4.3%) in exons 19 (L747_P753>Q) or 21 (L858R and L861Q) as determined by pyrosequencing. However, Cobas and PNA-clamping failed to detect the presence of EGFR gene mutation in these five cases.
CONCLUSION
No activating mutation of EGFR gene of clinical significance was observed in 148 TNBC cases using three commercially available methods. Thus, EGFR gene mutation appears to be an extremely rare event in patients with TNBC.

Keyword

Breast neoplasms; Epidermal growth factor receptor; Mutation; Triple negative breast neoplasms

MeSH Terms

Antibodies
Breast Neoplasms
Disease-Free Survival
Epidermal Growth Factor*
Exons
Genes, erbB-1
Humans
Immunohistochemistry
Incidence
Limit of Detection
Mutation Rate
Polymerase Chain Reaction
Receptor, Epidermal Growth Factor*
Triple Negative Breast Neoplasms*
Antibodies
Epidermal Growth Factor
Receptor, Epidermal Growth Factor

Figure

• 

  Figure 1 Immunohistochemical results for epidermal growth factor receptor (EGFR) protein as determined using pharmDx (2-18C9). (A) No staining or faint membranous staining in <10% of tumor cells (score 0), (B) faint membranous staining in ≥10% of tumor cells (score 1+), (C) moderate staining in ≥10% of tumor cells (score 2+), (D) strong membranous staining of ≥10% of tumor cells (score 3+) (original magnification ×400, in all figures).

• 

  Figure 2 Kaplan-Meier survival curves for epidermal growth factor receptor (EGFR) protein expression. (A) Overall survival, (B) disease-free survival.

• 

  Figure 3 Representative pyrosequencing results for EGFR gene mutations. (A) Case with 10.9% mutation frequency for the L858R mutation in EGFR exon 21. (B) Case with 9.0% mutation frequency for the L861Q mutation in EGFR exon 21. (C) Case with 10% mutation frequency for exon 19 deletion, L747_P753>Q. The numbers on the Y axis represent relative fluorescence unit. EGFR=epidermal growth factor receptor; LOD=limit of detection.

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