Related Experiment Video
Updated: May 15, 2026
Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
Published on: October 8, 2015
Effect of dabrafenib on melanoma cell lines harbouring the BRAF(V600D/R) mutations
Giusy Gentilcore1, Gabriele Madonna, Nicola Mozzillo
1Department Melanoma, Istituto Nazionale Tumori Fondazione Pascale, Naples, Italy.
Background:
Conventional therapeutic agents are largely unsatisfactory into the treatment of malignant melanoma. Recently, an innovative approach based on inhibitors of the mutated BRAF gene (which represents the most prevalent alteration in melanoma patients) appears very promising from the clinical point of view. On this regard, a new compound, dabrafenib (GSK2118436), has been demonstrated to be effective in patients carrying the BRAFV600E/K mutations. We here tested dabrafenib for its capability to inhibit cell growth on primary melanoma cell lines, established from patients' tumour tissues and carrying the BRAFV600D/R mutations.
Methods:
Three melanoma cell lines were tested: M257 wild-type BRAF, LCP BRAFV600R and WM266 BRAFV600D. The MTT assays were performed using standardized approaches. To evaluate the inhibition of MAPK pathway and the consequent inhibition of cellular proliferation, the phosphorylation of ERK was examined by Western Blot analysis performed on total protein extracts from cell lines after treatment with dabrafenib.
Results:
Our experiments demonstrated an effective action of Dabrafenib (GSK2118436) and the inhibition of MAPK pathway in melanoma cell lines carrying BRAFV600D/R mutations.
Conclusion:
These results could be helpful to enlarge the number of melanoma patients who may benefit of a more effective targeted treatment.
Insights
Dabrafenib effectively inhibits melanoma cell growth and the MAPK pathway in cell lines with BRAFV600D/R mutations. This targeted therapy shows promise for expanding treatment options for melanoma patients.
Area of Science:
- Oncology
- Molecular Biology
- Dermatology
Background:
- Malignant melanoma treatment remains challenging with conventional therapies.
- Targeted therapy using BRAF gene inhibitors offers a promising approach for melanoma.
- Dabrafenib has shown efficacy in patients with BRAFV600E/K mutations.
Purpose of the Study:
- To evaluate dabrafenib's efficacy in inhibiting cell growth.
- To assess dabrafenib's effect on melanoma cell lines with BRAFV600D/R mutations.
Main Methods:
- Tested dabrafenib on three melanoma cell lines (M257, LCP, WM266) with varying BRAF mutations.
- Utilized MTT assays to measure cell viability and proliferation.
- Examined MAPK pathway inhibition via Western Blot analysis of ERK phosphorylation.
Main Results:
- Dabrafenib demonstrated effective inhibition of cell growth in melanoma cell lines.
- The drug successfully inhibited the MAPK pathway in cell lines with BRAFV600D/R mutations.
Conclusions:
- Dabrafenib is effective against melanoma cell lines harboring BRAFV600D/R mutations.
- These findings may expand the patient population eligible for targeted BRAF inhibitor therapy.
Related Concept Videos
01:07MAPK Signaling Cascades
02:38Mitogens and the Cell Cycle