Journal: Cancer Cell International

Article Title: PHI-501, a dual inhibitor of RAF and DDR1/2, overcomes MAPK drug resistance in Melanoma

doi: 10.1186/s12935-026-04271-w

Figure Lengend Snippet: MAPK inhibitor-resistant melanoma cells exhibit enhanced DDR1/2 signaling. ( A ) Clonogenic assay showing the validation of drug-resistant cell lines. ( B ) Quantification of clonogenic assay shown in Fig. 1A. Data are presented as relative colony formation (%) normalized to the control group and expressed as the mean ± SEM, n = 3 replicates. Statistical significance was determined by two-way ANOVA. * p < 0.05, # p < 0.001, † p < 0.001, ‡ p < 0.0001. ( C ) Cell viability assay of RAF and MEK inhibitors in parental cells and drug-resistant cells for 48 h. Data are presented as the mean ± SEM, n = 3 replicates. ( D ) Western blot analysis of pERK and pAKT signaling responses in parental and resistant melanoma cells following 48-h treatment with vehicle or the corresponding resistance-selecting drug(s) at 1 µM. ( E ) Bubble plot representing Gene Set Enrichment Analysis (GSEA) using hallmark gene set comparing parental and resistant melanoma cells. Pathway enrichment was calculated using GSEA, with color indicating NES (blue = downregulated, red = upregulated) and bubble size representing −log10(p-value). ( F ) SK-MEL-2 and SK-MEL-2BR cells were stimulated with type I collagen (40 µg/ml) for 18 h to activate DDR signaling

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies diluted in 5% BSA in TBST as indicated in the text: p-DDR1 (Cell Signaling Technology, CST14531 ), p-DDR1/2 (R&D Systems, MAB25382), p-AKT (Cell Signaling Technology, CST4058), AKT (Cell Signaling Technology, CST9272), p-MEK (Cell Signaling Technology, CST9121), p-ERK (Cell Signaling Technology, CST9101), ERK (Cell Signaling Technology, CST9102), Cyclin D1(Cell Signaling Technology, CST2978), and Survivin (Cell Signaling Technology, CST2808).

Techniques: Clonogenic Assay, Biomarker Discovery, Control, Viability Assay, Western Blot

Journal: Cancer Cell International

Article Title: PHI-501, a dual inhibitor of RAF and DDR1/2, overcomes MAPK drug resistance in Melanoma

doi: 10.1186/s12935-026-04271-w

Figure Lengend Snippet: DDR1 and DDR2 are associated with MAPK pathway activation and therapeutic resistance in melanoma. ( A ) Kaplan–Meier survival analysis showing patient outcomes based on DDR1 and DDR2 expression levels across all TCGA cancer types. Patients were stratified into high (top 25%) and low (bottom 25%) expression groups, and overall survival (OS) was analyzed. ( B ) Proteomic analysis of BRAF/NRAS-mutant SKCM. Box plots demonstrate differential expression of AKT and phosphorylated MEK1 (pMEK1-S217/S221) between wild-type and mutant groups. ( C ) Correlation heatmaps between DDR1/2 and key genes in the MAPK and AKT signaling pathways in TCGA-SKCM samples. Color intensity represents the correlation value, with red indicating a positive correlation. Significance levels are indicated by asterisks (* p < 0.05, ** p < 0.01, *** p < 0.001). ( D ) Correlation analysis revealing DDR1 expression (DepMap) and MAPK pathway inhibitor sensitivity (GDSC2) in melanoma cell lines. Scatter plots depicting correlations between DDR1 expression levels and IC50 to Dabrafenib (left) and Trametinib (right)

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies diluted in 5% BSA in TBST as indicated in the text: p-DDR1 (Cell Signaling Technology, CST14531 ), p-DDR1/2 (R&D Systems, MAB25382), p-AKT (Cell Signaling Technology, CST4058), AKT (Cell Signaling Technology, CST9272), p-MEK (Cell Signaling Technology, CST9121), p-ERK (Cell Signaling Technology, CST9101), ERK (Cell Signaling Technology, CST9102), Cyclin D1(Cell Signaling Technology, CST2978), and Survivin (Cell Signaling Technology, CST2808).

Techniques: Activation Assay, Expressing, Mutagenesis, Quantitative Proteomics, Protein-Protein interactions