engleski

BRAF gene mutations in malignant melanoma in Rijeka county

Introduction: B-Raf serin/threonin kinase, encoded by the BRAF gene, plays a role in regulating the MAP/ERK signaling pathway, which affects cell division and differentiation se. BRAF V600 mutation is correlated with better response to B-Raf inhibitors in patients with metastatic melanoma. Objective: Here we present the BRAF gene mutation analysis data in Rijeka County over the period of two years. Method: BRAF V600 mutation analysis was indicated for patients diagnosed with metastatic melanoma in Rijeka County. Formalin fixed paraffin embedded tissue was used for the analysis. Area selected by the pathologist containing more than 50% melanoma cells was used for DNA extraction (NucleoSpin Tissue kit, Macherey-Nagel, Germany). BRAF gene exon 15 was amplified using the primers and PCR conditions from Lazar et al. 2009. Mutation identification was performed using Sanger sequencing with afore mentioned primers (BigDye*Terminator v1.1, ABIprism310 genetic analyzer, Applied Biosystems, USA) Results: In total we analyzed BRAF mutation status for 19 patients. Seven patients (38%) were wild type for BRAF mutations. 10 patients (52%) presented the most common BRAF V600E mutation, one patient (5%) presented V600K mutation while one patient(5%) exhibited a very rare BRAF L597S mutation. Conclusion: BRAF V600 activating mutation is occurring in 40-70 % of malignant melanoma depending on the melanoma subtype. Our data showed 58% of V600 mutants in analyzed melanomas. BRAF L597S mutation detected in one of our patients correlated with a significant response to a new therapy which has yet to be approved by the FDA (Dahlman BK et al. 2013). Sanger sequencing mutation analysis is not limited to already known mutations but rather detects all the mutations in the amplified region. Such data can be valuable in patient therapy and prospective studies.

metastatic melanoma; BRAF; Sanger sequencing

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