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Lee, Kim, Kim, Yoo, Kim, Oh, and Kim: PNA-Mediated PCR Clamping for the Detection of EGFR Mutations in Non-Small Cell Lung Cancer
Original Article
Tuberculosis and Respiratory Diseases

Tuberculosis and Respiratory Diseases 2010; 69(4): 271-278.

Published online: 31 October 2010

DOI: https://doi.org/10.4046/trd.2010.69.4.271

PNA-Mediated PCR Clamping for the Detection of EGFR Mutations in Non-Small Cell Lung Cancer

Kye Young Lee, M.D., Ph.D.1, Hee Joung Kim, M.D.1, Sun Jong Kim, M.D.1, Gwang Ha Yoo, M.D.1, Won Dong Kim, M.D.1, Seo Young Oh, M.S.2, Wan Seop Kim, M.D.2

1Department of Internal Medicine, Konkuk University School of Medicine, Seoul, Korea.

2Department of Pathology, Konkuk University School of Medicine, Seoul, Korea.

Address for correspondence: Kye Young Lee, M.D., Ph.D. Department of Internal Medicine, Konkuk University School of Medicine, 4-12, Hwayang-dong, Gwangjin-gu, Seoul 143-729, Korea. Phone: 82-2-2030-7521, Fax: 82-2-2030-7748, kyleemd@kuh.ac.kr

Received 3 September 2010       Accepted 15 October 2010

Copyright © 2010. The Korean Academy of Tuberculosis and Respiratory Diseases. All rights reserved.

Abstract

Background

Recent studies have demonstrated that the epidermal growth factor receptor (EGFR) genotype is the most important predictive marker to EGFR-tyrosine kinase inhibitors (TKIs) and first-line gefitinib treatment will be approved in the near future for use in non-small cell lung cancer (NSCLC) patients with the EGFR mutation. Direct sequencing is known to be the standard for detecting EGFR mutations; however, it has limited sensitivity. Peptide nucleic acids (PNA)-mediated PCR clamping method is a newly introduced method for analyzing EGFR mutations with increased sensitivity and stability.

Methods

A total of 71 NSCLC patients were analyzed for EGFR mutations using the PNA-mediated PCR clamping technique. Sixty-nine patients were analyzed for clinicopathologic correlation with EGFR genotype; 2 patients with indeterminate results were excluded. In order to determine EGFR-TKI drug response, 57 patients (42 gefitinib, 15 erlotinib) were included in the analysis.

Results

The EGFR mutation rate was 47.8%. Being female, a non-smoker, and having adenocarcinoma were favorable clinicopathologic factors, as expected. However, more than a few smokers (33.3%), male (28.1%), and patients with non-adenocarcinoma (28.6%) had the EGFR mutation. Having a combination of favorable clinicopathologic factors did not increase the EGFR mutation rate significantly. Drug response to EGFR-TKIs showed significant differences depending on the EGFR genotype; ORR was 14.3% for wild type vs 69.0% for mutant type; DCR is 28.6% for wild type vs 96.6% for mutant type. The median EGFR-TKI treatment duration is 7.6 months for mutant type group and 1.4 months for wild type group.

Conclusion

EGFR genotype determined using the PNA-mediated PCR clamping method is significantly correlated with the clinical EGFR-TKI responses and PNA-mediated PCR.

Keywords: Peptide Nucleic Acids, Receptor, Epidermal Growth Factor, Carcinoma, Non-Small-Cell Lung

Figures and Tables

Figure 1
The proportions of favorable clinicopathologic factors and each combination in EGFR mutation positive group (n=33).
trd-69-271-g001
Figure 2
Progression free survival (duration of EGFR-TKI treatment) according to EGFR genotype.
trd-69-271-g002
Table 1
Patient characteristics (n=71)
trd-69-271-i001
Table 2
The PCR protocol
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Table 3
The PNA-mediated clamping method detects 29 mutations of the EGFR gene
trd-69-271-i003

PNA: peptide nucleic acids; EGFR: epidermal growth factor receptor.

Table 4
EGFR mutation testing results using PNA-mediated PCR clamping (n=71)
trd-69-271-i004

Values are presented as number (%).

EGFR: epidermal growth factor receptor; PNA: peptide nucleic acids.

Table 5
Comparisons of clinicopathologic factors depending on EGFR genotype
trd-69-271-i005

EGFR: epidermal growth factor receptor.

Table 6
EGFR mutation positive rate depending on favorable clinicopathologic factors
trd-69-271-i006

EGFR: epidermal growth factor receptor.

Table 7
The concordance rate between EGFR genotypes and TKI responses
trd-69-271-i007

EGFR: epidermal growth factor receptor; TKI: tyrosine kinase inhibitors.

Acknowledgements

This study was supported by a grant from Korean Association for the Study of Lung Cancer (KASLC-1002).

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