Detection of EGFR and BRAF mutations by competitive allele-specific TaqMan polymerase chain reaction in lung adenocarcinoma

Yang,Yang; Meng,Yi; Zhang,Hang; Shen,Xiaoyan; Li,Rutian; Yu,Lixia; Liu,Baorui; Wang,Lifeng

doi:10.3892/ol.2017.7652

Detection of EGFR and BRAF mutations by competitive allele-specific TaqMan polymerase chain reaction in lung adenocarcinoma

Authors:
- Yang Yang
- Yi Meng
- Hang Zhang
- Xiaoyan Shen
- Rutian Li
- Lixia Yu
- Baorui Liu
- Lifeng Wang
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Affiliations: Department of Oncology, The Affiliated Taikang Xianlin Drum Tower Hospital of Medical School of Nanjing University, Nanjing, Jiangsu 210046, P.R. China, The Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University and Clinical Cancer Institute of Nanjing University, Nanjing, Jiangsu 210008, P.R. China
Published online on: December 19, 2017 https://doi.org/10.3892/ol.2017.7652
Pages: 3295-3304

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Abstract

Epithelial growth factor receptor (EGFR)-tyrosine kinase inhibitors are the standard first‑line treatment for patients with metastatic non‑small cell lung cancer (NSCLC) expressing sensitive EGFR‑mutants. Other drugs target different driver mutants, including the serine/threonine‑protein kinase B‑raf (BRAF) inhibitor dabrafenib, which has exhibited promising efficacy for treating patients with metastatic BRAF‑mutated NSCLC. Therefore, identifying patients carrying mutations that may be treated using targeted therapies is important. However, the methods of molecular detection presently applied in clinical practice, particularly detection of BRAF in NSCLC patients, require further investigation. Therefore, more sensitive and economic methods are required. The present study applied the competitive allele‑specific TaqMan polymerase chain reaction (CastPCR) technology to the molecular detection of EGFR (del2235‑2249, del2236‑2250, T790M, L858R) and BRAF (V600E, G469A, D594G) mutations in 144 treatment‑naive patients with lung adenocarcinoma, and analyzed the association between the mutation rates and patients' clinicopathological features. 51.4% (74/144) cases were identified harboring EGFR mutations. A total of 40.3% (58/144) patients carried sensitizing mutations (exon 19 deletion or L858R) and 14.6% (21/144) carried T790M mutations. 6.9% (10/144) mutation‑positive patients were double‑mutated. Total EGFR mutation rate was significantly increased in female compared with that of males (60.9 vs. 43.8%, P<0.05), in non‑smokers compared with that of smokers (62.8 vs. 34.5%, P<0.05). In total, 8.3% (12/144) patients were identified with BRAF mutations. 16.7% were V600E (2/12) and 83.3% (10/12) were non‑V600E mutants. Among the 10 non‑V600E mutations, D594G accounted for 90.0% (9/10) and G469A accounted for 10.0% (1/10). Statistical analysis demonstrated that the BRAF mutation rate was not associated with any of the following clinicopathological features: Sex, age, smoking history, clinical stages, distant metastasis, differentiation degree, tumor size and regional lymph node metastasis (P≥0.05). CastPCR technology is a robust method with high sensitivity for the molecular detection of EGFR and BRAF mutations in clinical formalin‑fixed paraffin-embedded samples.

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