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Rapid detection of epidermal growth factor receptor mutations with multiplex PCR and primer extension in lung cancer

Abstract Epidermal growth factor receptor (EGFR) kinase domain mutations hyperactivate the kinase and confer kinase addiction of the non-small-cell lung cancer (NSCLC) tumor cells. Almost all of these mutations are located within exons 18-21. The -216 single nucleotide polymorphism in the promoter region is associated with increased EGFR production. We present a method for detecting these common mutations in 81 cases of NSCLC. The protocol is based on the multiplex amplification of promoter region and exons 18-21 of the EGFR genes in a single tube, followed by primer extension of the PCR products using various sizes of primers to detect. | Lin et al. Journal of Biomedical Science 2010 17 37 http www.jbiomedsci.eom content 17 1 37 a NSC The cost of publication In Journal of Blomodlcal Science Is bome by the National Science Council Taiwan JOURNAL OF BIOMEDICAL SCIENCE RESEARCH Open Access Rapid detection of epidermal growth factor receptor mutations with multiplex PCR and primer extension in lung cancer Ching-Hsiung Lin1 Kun-Tu Yeh2 Ya-Sian Chang3 4 Nicholas C Hsu3 and Jan-Gowth Chang 3 5 6 Abstract Epidermal growth factor receptor EGFR kinase domain mutations hyperactivate the kinase and confer kinase addiction of the non-small-cell lung cancer NSCLC tumor cells. Almost all of these mutations are located within exons 18-21. The -216 single nucleotide polymorphism in the promoter region is associated with increased EGFR production. We present a method for detecting these common mutations in 81 cases of NSCLC. The protocol is based on the multiplex amplification of promoter region and exons 18-21 of the EGFR genes in a single tube followed by primer extension of the PCR products using various sizes of primers to detect base changes at -216 promoter region and codons 719 746750 790 858 of the EGFR gene. We compared the results with that from direct sequencing for detecting EGFR mutations in 81 cases of NSCLC. The two methods identified the same 26 mutations but our method is superior to direct sequencing in terms of the amount of work and time required. We presented a simple and fast method to detect mutations of EGFR genes in NSCLC. Background Lung cancer is one of the most common cancers in the world and is responsible for one third of all cancer-related death. Treatment of lung cancer mainly depends on the type of the cells that make up the cancer. Small-cell lung cancer SCLC which comprises about 20 of lung cancers originates from neuroendocrine cells in the bronchus. SCLC responds well to chemotherapy initially but resistance occurs commonly. Non-small-cell lung cancer NSCLC comprising 80 of lung .