Detection of EGFR mutations in circulating free DNA by PNA-mediated PCR clamping

Hye Ryoun Kim, Sung Yong Lee, Dae Sung Hyun, Min Ki Lee, Hyun Kyung Lee, Chang Min Choi, Sei Hoon Yang, Young Chul Kim, Yong Chul Lee, Sun Young Kim, Seung Hun Jang, Jae Cheol Lee, Kye Young Lee

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Background: Epidermal growth factor receptor (EGFR)-activating mutations are major determinants in predicting the tumor response to EGFR tyrosine kinase inhibitors in non-small cell lung cancer (NSCLC). Noninvasive test for the detection of EGFR mutations is required, especially in NSCLC patients from whom tissue is not available. In this study, we assessed the feasibility of detection of EGFR mutations in free DNA circulating in plasma. Methods. Plasma samples of 60 patients with partial response to gefitinib were analyzed to detect EGFR-activating mutations in exons 19 and 21. Forty (66.7%) of patients had tumor EGFR mutation results. EGFR mutations in plasma were detected using the peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method. All clinical data and plasma samples were obtained from 11 centers of the Korean Molecular Lung Cancer Group (KMLCG). Results: Of the 60 patients, 39 were female and the median age was 62.5 years. Forty-three patients never smoked, 53 had adenocarcinomas, and seven had other histologic types. EGFR-activating mutation was detected in plasma of 10 cases (exon 19 deletion in seven and exon 21 L858R point mutation in three). It could not be found in plasma after treatment for 2 months. When only patients with confirmed EGFR mutation in tumor were analyzed, 17% (6 of 35) of them showed positive plasma EGFR mutation and the mutation type was completely matched with that in tumor. There was no statistically significant difference in clinical parameters between patients with EGFR mutations in plasma and those without EGFR mutations. Conclusions: The detection rate of EGFR mutations from plasma was not so high despite highly sensitive EGFR mutation test suggesting that more advances in detection methods and further exploration of characteristics of circulating free DNA are required.

Original languageEnglish
Article number50
JournalJournal of Experimental and Clinical Cancer Research
Volume32
Issue number1
DOIs
Publication statusPublished - 2013 Aug 13

Fingerprint

Peptide Nucleic Acids
Epidermal Growth Factor Receptor
Constriction
Polymerase Chain Reaction
Mutation
DNA
Exons
Non-Small Cell Lung Carcinoma
Neoplasms
Point Mutation

Keywords

  • EGFR mutation
  • Non-small cell lung cancer
  • Plasma
  • PNA-mediated PCR clamping method

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Detection of EGFR mutations in circulating free DNA by PNA-mediated PCR clamping. / Kim, Hye Ryoun; Lee, Sung Yong; Hyun, Dae Sung; Lee, Min Ki; Lee, Hyun Kyung; Choi, Chang Min; Yang, Sei Hoon; Kim, Young Chul; Lee, Yong Chul; Kim, Sun Young; Jang, Seung Hun; Lee, Jae Cheol; Lee, Kye Young.

In: Journal of Experimental and Clinical Cancer Research, Vol. 32, No. 1, 50, 13.08.2013.

Research output: Contribution to journalArticle

Kim, HR, Lee, SY, Hyun, DS, Lee, MK, Lee, HK, Choi, CM, Yang, SH, Kim, YC, Lee, YC, Kim, SY, Jang, SH, Lee, JC & Lee, KY 2013, 'Detection of EGFR mutations in circulating free DNA by PNA-mediated PCR clamping', Journal of Experimental and Clinical Cancer Research, vol. 32, no. 1, 50. https://doi.org/10.1186/1756-9966-32-50
Kim, Hye Ryoun ; Lee, Sung Yong ; Hyun, Dae Sung ; Lee, Min Ki ; Lee, Hyun Kyung ; Choi, Chang Min ; Yang, Sei Hoon ; Kim, Young Chul ; Lee, Yong Chul ; Kim, Sun Young ; Jang, Seung Hun ; Lee, Jae Cheol ; Lee, Kye Young. / Detection of EGFR mutations in circulating free DNA by PNA-mediated PCR clamping. In: Journal of Experimental and Clinical Cancer Research. 2013 ; Vol. 32, No. 1.
@article{88a6bb3b68bb4bfeaeb07521b0725233,
title = "Detection of EGFR mutations in circulating free DNA by PNA-mediated PCR clamping",
abstract = "Background: Epidermal growth factor receptor (EGFR)-activating mutations are major determinants in predicting the tumor response to EGFR tyrosine kinase inhibitors in non-small cell lung cancer (NSCLC). Noninvasive test for the detection of EGFR mutations is required, especially in NSCLC patients from whom tissue is not available. In this study, we assessed the feasibility of detection of EGFR mutations in free DNA circulating in plasma. Methods. Plasma samples of 60 patients with partial response to gefitinib were analyzed to detect EGFR-activating mutations in exons 19 and 21. Forty (66.7{\%}) of patients had tumor EGFR mutation results. EGFR mutations in plasma were detected using the peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method. All clinical data and plasma samples were obtained from 11 centers of the Korean Molecular Lung Cancer Group (KMLCG). Results: Of the 60 patients, 39 were female and the median age was 62.5 years. Forty-three patients never smoked, 53 had adenocarcinomas, and seven had other histologic types. EGFR-activating mutation was detected in plasma of 10 cases (exon 19 deletion in seven and exon 21 L858R point mutation in three). It could not be found in plasma after treatment for 2 months. When only patients with confirmed EGFR mutation in tumor were analyzed, 17{\%} (6 of 35) of them showed positive plasma EGFR mutation and the mutation type was completely matched with that in tumor. There was no statistically significant difference in clinical parameters between patients with EGFR mutations in plasma and those without EGFR mutations. Conclusions: The detection rate of EGFR mutations from plasma was not so high despite highly sensitive EGFR mutation test suggesting that more advances in detection methods and further exploration of characteristics of circulating free DNA are required.",
keywords = "EGFR mutation, Non-small cell lung cancer, Plasma, PNA-mediated PCR clamping method",
author = "Kim, {Hye Ryoun} and Lee, {Sung Yong} and Hyun, {Dae Sung} and Lee, {Min Ki} and Lee, {Hyun Kyung} and Choi, {Chang Min} and Yang, {Sei Hoon} and Kim, {Young Chul} and Lee, {Yong Chul} and Kim, {Sun Young} and Jang, {Seung Hun} and Lee, {Jae Cheol} and Lee, {Kye Young}",
year = "2013",
month = "8",
day = "13",
doi = "10.1186/1756-9966-32-50",
language = "English",
volume = "32",
journal = "Journal of Experimental and Clinical Cancer Research",
issn = "0392-9078",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Detection of EGFR mutations in circulating free DNA by PNA-mediated PCR clamping

AU - Kim, Hye Ryoun

AU - Lee, Sung Yong

AU - Hyun, Dae Sung

AU - Lee, Min Ki

AU - Lee, Hyun Kyung

AU - Choi, Chang Min

AU - Yang, Sei Hoon

AU - Kim, Young Chul

AU - Lee, Yong Chul

AU - Kim, Sun Young

AU - Jang, Seung Hun

AU - Lee, Jae Cheol

AU - Lee, Kye Young

PY - 2013/8/13

Y1 - 2013/8/13

N2 - Background: Epidermal growth factor receptor (EGFR)-activating mutations are major determinants in predicting the tumor response to EGFR tyrosine kinase inhibitors in non-small cell lung cancer (NSCLC). Noninvasive test for the detection of EGFR mutations is required, especially in NSCLC patients from whom tissue is not available. In this study, we assessed the feasibility of detection of EGFR mutations in free DNA circulating in plasma. Methods. Plasma samples of 60 patients with partial response to gefitinib were analyzed to detect EGFR-activating mutations in exons 19 and 21. Forty (66.7%) of patients had tumor EGFR mutation results. EGFR mutations in plasma were detected using the peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method. All clinical data and plasma samples were obtained from 11 centers of the Korean Molecular Lung Cancer Group (KMLCG). Results: Of the 60 patients, 39 were female and the median age was 62.5 years. Forty-three patients never smoked, 53 had adenocarcinomas, and seven had other histologic types. EGFR-activating mutation was detected in plasma of 10 cases (exon 19 deletion in seven and exon 21 L858R point mutation in three). It could not be found in plasma after treatment for 2 months. When only patients with confirmed EGFR mutation in tumor were analyzed, 17% (6 of 35) of them showed positive plasma EGFR mutation and the mutation type was completely matched with that in tumor. There was no statistically significant difference in clinical parameters between patients with EGFR mutations in plasma and those without EGFR mutations. Conclusions: The detection rate of EGFR mutations from plasma was not so high despite highly sensitive EGFR mutation test suggesting that more advances in detection methods and further exploration of characteristics of circulating free DNA are required.

AB - Background: Epidermal growth factor receptor (EGFR)-activating mutations are major determinants in predicting the tumor response to EGFR tyrosine kinase inhibitors in non-small cell lung cancer (NSCLC). Noninvasive test for the detection of EGFR mutations is required, especially in NSCLC patients from whom tissue is not available. In this study, we assessed the feasibility of detection of EGFR mutations in free DNA circulating in plasma. Methods. Plasma samples of 60 patients with partial response to gefitinib were analyzed to detect EGFR-activating mutations in exons 19 and 21. Forty (66.7%) of patients had tumor EGFR mutation results. EGFR mutations in plasma were detected using the peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method. All clinical data and plasma samples were obtained from 11 centers of the Korean Molecular Lung Cancer Group (KMLCG). Results: Of the 60 patients, 39 were female and the median age was 62.5 years. Forty-three patients never smoked, 53 had adenocarcinomas, and seven had other histologic types. EGFR-activating mutation was detected in plasma of 10 cases (exon 19 deletion in seven and exon 21 L858R point mutation in three). It could not be found in plasma after treatment for 2 months. When only patients with confirmed EGFR mutation in tumor were analyzed, 17% (6 of 35) of them showed positive plasma EGFR mutation and the mutation type was completely matched with that in tumor. There was no statistically significant difference in clinical parameters between patients with EGFR mutations in plasma and those without EGFR mutations. Conclusions: The detection rate of EGFR mutations from plasma was not so high despite highly sensitive EGFR mutation test suggesting that more advances in detection methods and further exploration of characteristics of circulating free DNA are required.

KW - EGFR mutation

KW - Non-small cell lung cancer

KW - Plasma

KW - PNA-mediated PCR clamping method

UR - http://www.scopus.com/inward/record.url?scp=84881137132&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84881137132&partnerID=8YFLogxK

U2 - 10.1186/1756-9966-32-50

DO - 10.1186/1756-9966-32-50

M3 - Article

C2 - 23927790

AN - SCOPUS:84881137132

VL - 32

JO - Journal of Experimental and Clinical Cancer Research

JF - Journal of Experimental and Clinical Cancer Research

SN - 0392-9078

IS - 1

M1 - 50

ER -