Capillary electrophoretic genotyping of epidermal growth factor receptor for pharmacogenomic assay of lung cancer therapy.
In this research, a universal multiplex PCR method combining capillary gel electrophoresis was established for simultaneous analysis of mutations in exons 18-21 of EGFR kinase domain gene. After rinsing of the gel solution (1 × TBE buffer containing 2% HEC and 1.5% HPC), the diluted PCR sample was electrokinetically injected and separated at 35°C. Several parameters were investigated for method optimization, including the kinds of polymer, capillary temperature and separation voltage. Under optimal conditions, time taken for analyzing mutations of EGFR kinase domain gene in lung cancer patients was short (within 15 min). A total of 50 lung cancer patients' tumor tissues were analyzed, and two frequent mutations associated with therapeutic efficiency of lung cancer were identified, including deletion of exon 19 (8/50) and L858R point mutation in exon 21 (12/50). This method is feasible for application in pharmacogenomic assay of lung cancer therapy. By applying this simple and effective method for genotyping of lung cancer patients, the information of pharmacogenomics was supplied for lung cancer therapy and to assist diagnosis to prolong patients' lives.
School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan.
Chun-Chi
CC
Wang
Kun-Hsien
KH
Chao
Yen-Ling
YL
Chen
Jan-Gowth
JG
Chang
Shou-Mei
SM
Wu
D001483:Base Sequence; D017931:DNA Primers; D019075:Electrophoresis, Capillary / Q000379:methods*; D066246:ErbB Receptors / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000628:therapy*; D010597:Pharmacogenetics; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D066246:ErbB Receptors