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Modified Restriction Enzyme-based Detection of EGFR Mutations in Lung Cancer

Author(s): Fan YANG | Kezhong CHEN | Guanchao JIANG | Jianfeng LI | Jun WANG

Journal: Chinese Journal of Lung Cancer
ISSN 1009-3419

Volume: 14;
Issue: 8;
Start page: 637;
Date: 2011;
Original page

Keywords: Lung neoplasms | Epidermal growth factor receptor | Mutation

Background and objective Epidermal growth factor receptor (EGFR) gene mutation in lung cancer is a reliable predictor of the efficacy of tyrosine kinase inhibitors, so detection of gene mutation has very important clinical significance. The aim of this study is to optimize conditions for restriction enzyme-based mutant enriched detection of EGFR mutation in lung cancer specimens. Methods EGFR somatic mutations identified in DNA specimens from 251 lung adenocarcinomas with modified mutant enriched method and direct sequencing were compared. The sensitivity of the modified method was determined by serial dilution of mutant-carrying lung cancer cells in wild-type cells. Results 46 deletion mutations in exon 19 and 26 point mutations in exon 21 were found by sequencing, while modified method identified another 78 deletions in exon 19 and 57 substitutions in exon 21 with a mutation rate of 53.8%. Sensitivity of this modified method was 0.5% (mutant/wild type) in serial diluted cells. Conclusion The modified restriction enzyme-based method is convenient for clinical EGFR mutation screening in non-small cell lung cancer for its simpleness, cost-saving and high sensitivity.
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