Ho Chi Minh City Open University Journal of Science - Engineering and Technology (Feb 2015)
Egfr and K-ras in molecularly targeted therapy: From In Silico to In Vitro study
Abstract
Molecularly targeted therapy is a new type of cancer treatment which uses monoclonalantibodies or small substances to identify and attack cancer cells, except normal cell. This has more advantages than classical treatments. EGFR (Epidermal Growth Factor Receptor) and Kras (Kirsten-ras) are considered as two well molecular-targeted agent. In present study, we performed a systematic literature review in NCBI and computed average frequencies of EGFR and K-ras mutations in some common cancers. According to data analysis, we conclude that global published mutations which belong to these genes neither exclude each other nor associate with factors of race. We successfully designed and evaluated (1) primers for detection of EGFR and K-ras mutations by PCR-sequencing; (2) primer/probe sets are used for detection of the most seven common K-ras mutations by Allele-Specific-Realtime PCR (AS-Real-time PCR) as well as Allele-Specific PCR. Due to initial experimental results, an appropriate DNA extraction procedure from FFPE (Formalin-Fixed, ParaffinEmbedded) samples and optimal Tm for each primer couple of amplified mutation, containing regions on K-ras have been drawn. Results of K-ras mutation on colorectal cancer patients obtained from Tay Ninh Province, Viet Nam was firstly detected by using PCR-sequencing methods, then, confirmed by AS-Real-Time PCR.