The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer.
Lung cancer is the leading cause of cancer-related death worldwide. Recent progress in lung cancer diagnosis and treatment has been achieved due to a better understanding the molecular mechanisms of the disease and the identification of biomarkers that allow more specific cancer treatments. One of t...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0117983&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850023218476220416 |
|---|---|
| author | Marzena Anna Lewandowska Karol Czubak Katarzyna Klonowska Wojciech Jozwicki Janusz Kowalewski Piotr Kozlowski |
| author_facet | Marzena Anna Lewandowska Karol Czubak Katarzyna Klonowska Wojciech Jozwicki Janusz Kowalewski Piotr Kozlowski |
| author_sort | Marzena Anna Lewandowska |
| collection | DOAJ |
| description | Lung cancer is the leading cause of cancer-related death worldwide. Recent progress in lung cancer diagnosis and treatment has been achieved due to a better understanding the molecular mechanisms of the disease and the identification of biomarkers that allow more specific cancer treatments. One of the best known examples of personalized therapy is the use of tyrosine kinase inhibitors, such as gefitinib and erlotinib, for the successful treatment of non-small-cell lung cancer patients selected based on the specific EGFR mutations. Therefore, the reliable detection of mutations is critical for the application of appropriate therapy. In this study, we tested a two-tiered mutation detection strategy using real-time PCR assays as a well-validated high-sensitivity method and multiplex ligation-dependent probe amplification (MLPA)-based EGFRmut+ assay as a second-tier standard-sensitivity method. One additional advantage of the applied MLPA method is that it allows the simultaneous detection of EGFR mutations and copy-number alterations (i.e., amplifications) in EGFR, MET and ERBB2. Our analysis showed high concordance between these two methods. With the use of this two-tier strategy, we reliably determined the frequency of EGFR mutations and EGFR, MET and ERBB2 amplifications in over 200 lung cancer samples. Additionally, taking advantage of simultaneous copy number and small mutation analyses, we showed a very strong correlation between EGFR mutations and EGFR amplifications and a mutual exclusiveness of EGFR mutations/amplifications with MET and ERBB2 amplifications. Our results proved the reliability and usefulness of the two-tiered EGFR testing strategy. |
| format | Article |
| id | doaj-art-5a642e35faf44b4abacb5cdcb5c0af2e |
| institution | DOAJ |
| issn | 1932-6203 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-5a642e35faf44b4abacb5cdcb5c0af2e2025-08-20T03:01:27ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01102e011798310.1371/journal.pone.0117983The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer.Marzena Anna LewandowskaKarol CzubakKatarzyna KlonowskaWojciech JozwickiJanusz KowalewskiPiotr KozlowskiLung cancer is the leading cause of cancer-related death worldwide. Recent progress in lung cancer diagnosis and treatment has been achieved due to a better understanding the molecular mechanisms of the disease and the identification of biomarkers that allow more specific cancer treatments. One of the best known examples of personalized therapy is the use of tyrosine kinase inhibitors, such as gefitinib and erlotinib, for the successful treatment of non-small-cell lung cancer patients selected based on the specific EGFR mutations. Therefore, the reliable detection of mutations is critical for the application of appropriate therapy. In this study, we tested a two-tiered mutation detection strategy using real-time PCR assays as a well-validated high-sensitivity method and multiplex ligation-dependent probe amplification (MLPA)-based EGFRmut+ assay as a second-tier standard-sensitivity method. One additional advantage of the applied MLPA method is that it allows the simultaneous detection of EGFR mutations and copy-number alterations (i.e., amplifications) in EGFR, MET and ERBB2. Our analysis showed high concordance between these two methods. With the use of this two-tier strategy, we reliably determined the frequency of EGFR mutations and EGFR, MET and ERBB2 amplifications in over 200 lung cancer samples. Additionally, taking advantage of simultaneous copy number and small mutation analyses, we showed a very strong correlation between EGFR mutations and EGFR amplifications and a mutual exclusiveness of EGFR mutations/amplifications with MET and ERBB2 amplifications. Our results proved the reliability and usefulness of the two-tiered EGFR testing strategy.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0117983&type=printable |
| spellingShingle | Marzena Anna Lewandowska Karol Czubak Katarzyna Klonowska Wojciech Jozwicki Janusz Kowalewski Piotr Kozlowski The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer. PLoS ONE |
| title | The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer. |
| title_full | The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer. |
| title_fullStr | The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer. |
| title_full_unstemmed | The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer. |
| title_short | The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer. |
| title_sort | use of a two tiered testing strategy for the simultaneous detection of small egfr mutations and egfr amplification in lung cancer |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0117983&type=printable |
| work_keys_str_mv | AT marzenaannalewandowska theuseofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT karolczubak theuseofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT katarzynaklonowska theuseofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT wojciechjozwicki theuseofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT januszkowalewski theuseofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT piotrkozlowski theuseofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT marzenaannalewandowska useofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT karolczubak useofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT katarzynaklonowska useofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT wojciechjozwicki useofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT januszkowalewski useofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer AT piotrkozlowski useofatwotieredtestingstrategyforthesimultaneousdetectionofsmallegfrmutationsandegframplificationinlungcancer |