| Summary: | 碩士 === 高雄醫學大學 === 藥學研究所 === 98 === The observation of sensitivity to EGFR-TKIs (epidermal growth factor receptor - tyrosine kinase inhibitors), gefitinib and erlotinib, correlated very strongly with a newly discovered class of somatic activating mutations in the EGFR kinase domain. EGFR kinase domain mutations target four exons (18–21), which encoded part of the tyrosine kinase domain. EGFR-TKI is a type of medication that blocks the growth of cancer cells by interfering with specific targeted molecules needed for carcinogenesis and tumor growth, more effective than chemotherapy and less harmful to normal cells. EGFR genotyping in lung cancer plays an important role in decisions of treatment.
We have established a multiplex PCR method combining capillary gel electrophoresis for simultaneous analysis of EGFR kinase domain mutations. By optimized CE condition, the CE separation was performed in DB-17 capillary. Running buffer was 1x TBE buffer containing 2% HEC and 1.5% HPC；applied voltage was -10 kV and separation temperature was at 35℃. Under these optimal conditions, it took shorter time to analyze the polymorphism of EGFR kinase domain of lung cancer patients. This method is feasible for application in pharmacogenomics of lung cancer therapy. We performed the genotyping of 50 patients, and corresponded with DNA sequencing. By applying this simple and effective method, we can provide pharmacogenomics information of lung cancer therapy and assist diagnosis to prolong patients’ life.
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