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Detecting EGFR alterations in clinical specimens-pitfalls and necessities.

Isaksson, Sofi LU ; Bendahl, Pär-Ola LU ; Salomonsson, Annette LU ; Jönsson, Mats LU ; Haglund, Monica; Gaber, Alexander LU ; Jirström, Karin LU ; Jönsson, Per LU ; Borg, Åke LU and Johansson, Leif LU , et al. (2013) In Virchows Archiv: an international journal of pathology 463(6). p.755-764
Abstract
We investigated the epidermal growth factor receptor (EGFR) status in early stage lung cancer in Southern Sweden, a population for which there are no previous reports on the EGFR mutation frequency. Three hundred fifty small cell lung cancers, adenocarcinomas (AC), squamous cell carcinomas (SqCC), and large cell carcinomas were analyzed using a combination of techniques for the analysis of protein expression, gene copy numbers, and mutations. Immunohistochemical (IHC) staining with antibodies for the EGFR mutations L858R and del E746-A750 revealed intratumoral heterogeneity and several discrepant cases when compared to mutation-specific polymerase chain reaction (PCR)-based analysis. The frequencies of these two mutations, when considering... (More)
We investigated the epidermal growth factor receptor (EGFR) status in early stage lung cancer in Southern Sweden, a population for which there are no previous reports on the EGFR mutation frequency. Three hundred fifty small cell lung cancers, adenocarcinomas (AC), squamous cell carcinomas (SqCC), and large cell carcinomas were analyzed using a combination of techniques for the analysis of protein expression, gene copy numbers, and mutations. Immunohistochemical (IHC) staining with antibodies for the EGFR mutations L858R and del E746-A750 revealed intratumoral heterogeneity and several discrepant cases when compared to mutation-specific polymerase chain reaction (PCR)-based analysis. The frequencies of these two mutations, when considering IHC staining with mutation-specific antibodies in a cohort of 298 cases and subsequent confirmation by PCR, were 10 % in AC and <2 % in SqCC. Furthermore, screening by sequencing of EGFR in a cohort of 52 lung AC and squamous carcinomas demonstrated a more diverse mutation spectrum, not covered by the mutation-specific antibodies. High expression of total EGFR protein was correlated to high gene copy numbers but did not reflect the mutational status of the tumors. We believe that the mutation spectra in a Southern Swedish population is too diverse to be covered by the mutation-specific antibodies, and we also raise some other issues regarding the use of the mutation-specific antibodies, for example concerning heterogeneous expression of the mutated protein, optimal antibody dilution, and discrepancies between staining results and PCR. (Less)
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published
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Virchows Archiv: an international journal of pathology
volume
463
issue
6
pages
755 - 764
publisher
Springer
external identifiers
  • wos:000328214900004
  • pmid:24158511
  • scopus:84890549671
ISSN
1432-2307
DOI
10.1007/s00428-013-1489-y
language
English
LU publication?
yes
id
f49c9d6c-1fe3-40a9-8a27-0be2bcc5dc5b (old id 4143004)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/24158511?dopt=Abstract
date added to LUP
2013-11-04 22:16:58
date last changed
2019-03-13 17:33:20
@article{f49c9d6c-1fe3-40a9-8a27-0be2bcc5dc5b,
  abstract     = {We investigated the epidermal growth factor receptor (EGFR) status in early stage lung cancer in Southern Sweden, a population for which there are no previous reports on the EGFR mutation frequency. Three hundred fifty small cell lung cancers, adenocarcinomas (AC), squamous cell carcinomas (SqCC), and large cell carcinomas were analyzed using a combination of techniques for the analysis of protein expression, gene copy numbers, and mutations. Immunohistochemical (IHC) staining with antibodies for the EGFR mutations L858R and del E746-A750 revealed intratumoral heterogeneity and several discrepant cases when compared to mutation-specific polymerase chain reaction (PCR)-based analysis. The frequencies of these two mutations, when considering IHC staining with mutation-specific antibodies in a cohort of 298 cases and subsequent confirmation by PCR, were 10 % in AC and &lt;2 % in SqCC. Furthermore, screening by sequencing of EGFR in a cohort of 52 lung AC and squamous carcinomas demonstrated a more diverse mutation spectrum, not covered by the mutation-specific antibodies. High expression of total EGFR protein was correlated to high gene copy numbers but did not reflect the mutational status of the tumors. We believe that the mutation spectra in a Southern Swedish population is too diverse to be covered by the mutation-specific antibodies, and we also raise some other issues regarding the use of the mutation-specific antibodies, for example concerning heterogeneous expression of the mutated protein, optimal antibody dilution, and discrepancies between staining results and PCR.},
  author       = {Isaksson, Sofi and Bendahl, Pär-Ola and Salomonsson, Annette and Jönsson, Mats and Haglund, Monica and Gaber, Alexander and Jirström, Karin and Jönsson, Per and Borg, Åke and Johansson, Leif and Staaf, Johan and Planck, Maria},
  issn         = {1432-2307},
  language     = {eng},
  number       = {6},
  pages        = {755--764},
  publisher    = {Springer},
  series       = {Virchows Archiv:  an international journal of pathology},
  title        = {Detecting EGFR alterations in clinical specimens-pitfalls and necessities.},
  url          = {http://dx.doi.org/10.1007/s00428-013-1489-y},
  volume       = {463},
  year         = {2013},
}