Inhibiting insulin and mTOR signaling by afatinib and crizotinib combination fosters broad cytotoxic effects in cutaneous malignant melanoma

摘要

a Table showing loss in cell viability after single or combination treatment in 5 CMM cell lines (data presented as mean; n = 3 technical replicates; p < 0.001). b Volcano plots showing proteins altered in A375 (BRAF mutant) and SKMel2 (NRAS mutant) as assessed by LC MS-based phosphoproteomics or whole proteomics (WP) comparing combination treatment with DMSO, afatinib or crizotinib. Linear cut off p < 0.05 and 1.5 fold change (n = 3 technical replicates) was used to identify differentially expressed proteins, up-regulated (red) and down-regulated (green) proteins. Here we found that in phosphoproteomics the number of significantly differentially expressed proteins ranged between 300–1100 in the different treatment comparisons, whereas in WP it was around 400–1360 proteins. c Illustration of CMM cells and protein lysates extraction from A375 xenograft tumor tissue for RPPA. d Heat map with supervised clustering showing changes in selected candidate proteins analyzed by RPPA after 3 h or e 24 h of treatment with DMSO, single or combination drugs (n = 3).