Chinese medical journal, 2017-06, Vol.130 (12), p.1446-1453
2017
Details
- Autor(en) / Beteiligte
- Titel
- Establishment of a Novel Method for Screening Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Resistance Mutations in Lung Cancer
- Ist Teil von
- Chinese medical journal, 2017-06, Vol.130 (12), p.1446-1453
- Ort / Verlag
- China: Medknow Publications and Media Pvt. Ltd
- Erscheinungsjahr
- 2017
- Link zum Volltext
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- Background： Drug resistance to targeted therapies occurs in lung cancer, and resistance mechanisms related to epidermal growth factor receptor （EGFR） tyrosine kinase inhibitors （TKIs） are continuously being discovered. We aimed to establish a novel method for highly parallel multiplexed detection of genetic mutations related to EGFR TKl-resistant lung cancer using Agena iPLEX chemistry and niatrix-assisted laser desorption ionization time-of-flight analysis on the MassARRAY mass spectrometry platform. Methods： A review of the literature revealed 60 mutation hotspots in seven target genes （EGFR, KRAS, PIK3CA, BRAF, ERBB2, NRAS, and BIM） that are closely related to EGFR TKI resistance to lung cancer. A total of 183 primers comprised 61 paired forward and reverse amplification primers, and 61 matched extension primers were designed using Assay Design Software. The detection method was established by analyzing nine cell lines, and by comparison with LungCartaTM kit in ten lung cancer specimens. EGFR, KRAS. and BIM genes in all cell lines and clinical samples were subjected to Sanger sequencing for confirming reproducibility. Results： Our data showed that designed panel was a high-throughput and robust tool, allowing genotyping for sixty hotspots in the same run. Moreover, it made efficient use of patient diagnostic samples for a more accurate EGFR TKIs resistance analysis. The proposed method could accurately detect mutations in lung cancer cell lines and clinical specimens, consistent with those obtained by the LungCartaTM kit and Sanger sequencing. We also established a method for detection of large-fragment deletions based on single-base extension technology of MassARRAY platform. Conclusions： We established an effective method for high-throughput detection of genetic mutations related to EGFR TKI resistance based on the MassARRAY platform, which could provide more accurate information for overcoming cancers with de novo or acquired resistance to EGFR-targeted therapies.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0366-6999eISSN: 2542-5641DOI: 10.4103/0366-6999.207478Titel-ID: cdi_doaj_primary_oai_doaj_org_article_c14f5df97e7d4e04a6a541562b837a54
- Format
- –
- Schlagworte
- Analysis, Cell Line, Tumor, Design, Drug resistance, Drug Resistance, Neoplasm - drug effects, Drug Resistance, Neoplasm - genetics, Drug Resistance, Epidermal Growth Factor Receptor, Lung Cancer, MassARRAY, Targeted Molecular Therapy, Epidermal growth factor, Gene amplification, Gene mutations, Genetic Testing - methods, Genotype, Humans, Kinases, Lung cancer, Lung Neoplasms - enzymology, Lung Neoplasms - genetics, Medical research, Molecular weight, Mutation, Mutation - genetics, Oncology, Original, Protein Kinase Inhibitors - pharmacology, Receptor, Epidermal Growth Factor - antagonists & inhibitors, Receptor, Epidermal Growth Factor - genetics, Reproducibility of Results, Research centers, Tyrosine