Background
Materials and methods
Reagents and cell lines
BRAFV600E mutation analysis
Oncomap 3 core mass-spectrometric genotyping
SNP array analysis
Cell proliferation and viability assays
Phosphoflow staining
Cell cycle analysis
Apoptosis analysis
Western Blotting
In vitro metabolic tracer uptake assay
In vivo microCT and microPET studies
Statistical analysis
Results
PLX4032 specifically blocks the MAPK pathway in melanoma cell lines with the BRAFV600E mutation
Cell Line | NRAS/BRAF | Number of BRAF Gene Copies | Other Oncogenic Events | Cell line doubling time (hours) | PLX4032 IC50 (μM) |
---|---|---|---|---|---|
M257 | Wild type | 3 | CDKN2A R80 | 31.4 | Not reached |
M202 | NRAS Q61L | 2 | EGFR amplification CDKN2A homozygous deletion | 26.1 | Not reached |
M207 | NRAS Q61L | 2 | MITF amplification PTEN heterozygous deletion | 25.2 | Not reached |
M233 | BRAFV600E Heterozygous | 3 | AKT1 amplification CCND1 amplification EGFR amplification CDKN2A homozygous deletion PTEN homozygous deletion | 29.6 | Not reached |
M255 | BRAFV600E Heterozygous | 2 | AKT2 amplification CCND1 amplification EGFR amplification CDKN2A homozygous deletion | 48.6 | Not reached |
M308 | BRAFV600E Heterozygous | 3 | MITF amplification AKT2 amplification EGFR amplification CDKN2A heterozygous deletion | 35.0 | Not reached |
M263 | BRAFV600E Heterozygous | 2 | CDKN2A heterozygous deletion | 23.3 | 10 |
M321 | BRAFV600E Homozygous | 2 | 34.1 | 7.5 | |
SKMEL28 | BRAFV600E Homozygous | 2 | EGFR P753S MITF amplification CCND1 amplification CDKN2A heterozygous deletion PTEN heterozygous deletion | 26.9 | 4.6 |
M229 | BRAFV600E Homozygous | 4 | MITF amplification AKT1 amplification PTEN heterozygous deletion | 27.9 | 0.2 |
M238 | BRAFV600E Heterozygous | 2 | CDKN2A homozygous deletion PTEN heterozygous deletion | 28.1 | 0.7 |
M249 | BRAFV600E Heterozygous | 3 | MITF amplification AKT2 amplification PTEN homozygous deletion | 21.2 | 0.8 |
M262 | BRAFV600E Homozygous | 2 | AKT1 E17K AKT1 amplification EGFR amplification CDKN2A homozygous deletion | 47.4 | 0.1 |