A useful protocol for analyses of mutations of the epidermal growth factor receptor gene
| Autor: | Cheng-long Huang, Kouichi Yube, Dage Liu, Takashi Nakashima, Jun Nakano, Hiroyasu Yokomise, Masaki Ueno, Daiki Masuya |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Cancer Research
Lung Neoplasms Molecular Sequence Data Biology medicine.disease_cause Polymerase Chain Reaction chemistry.chemical_compound Growth factor receptor Carcinoma Non-Small-Cell Lung medicine Humans Epidermal growth factor receptor Gene Mutation Paraffin Embedding Base Sequence DNA Neoplasm Genes erbB-1 Sequence Analysis DNA General Medicine DNA extraction Oncology chemistry Cancer research biology.protein Nested polymerase chain reaction Tyrosine kinase DNA |
| Zdroj: | Oncology Reports. |
| ISSN: | 1791-2431 1021-335X |
| DOI: | 10.3892/or.15.6.1503 |
| Popis: | Recent studies have reported that mutations of the epidermal growth factor receptor (EGFR) gene are associated with the responsiveness of tyrosine kinase inhibitors (TKIs), which are molecular targets for non-small cell lung cancer (NSCLC). To provide genetic analyses for NSCLC patients, a simple and reliable method using paraffin-embedded materials is needed. The DEXPAT DNA extraction kit was used for DNA extraction from paraffin-embedded materials. DNA was amplified using the nested PCR technique, then analyzed by direct sequencing for EGFR mutations (exons 18 to 21). The phenol/chloroform extraction for DNA was also performed for comparison. When the DEXPAT kit was used, distinct bands were observed in all products after nested PCR assays of paraffin-embedded materials. Distinct sequencing signals were obtained. Results from the sequencing analysis of paraffin-embedded materials and frozen materials were completely concordant. The current study suggests that DNA extraction with the DEXPAT kit followed by nested PCR is a simple and reliable technique for analyzing the EGFR gene status with paraffin-embedded samples. |
| Databáze: | OpenAIRE |
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