Cc 115

The HNSCC cell lines CAL33, UD-SCC-2, UM-SCC-47, and HSC4 were provided by Dr. Thorsten Rieckmann (University Medical Center Hamburg-Eppendorf, Germany) and cultivated in Dulbecco’s MEM (DMEM; Gibco, Waltham, MA, USA) supplemented with 10% FBS (Merck, Darmstadt, Germany) and 1% penicillin/streptomycin (Gibco, Waltham, MA, USA). CAL33 and HSC4 are HPV⁻, while UM-SCC-47 and UD-SCC-2 are HPV⁺. SBLF7 and SBLF9 are normal human skin fibroblasts and were isolated from healthy donors by skin biopsy and subsequently cultivated with 15% FBS; 2% non-essential amino acids (NEA; Merck, Darmstadt, Germany); and 1% penicillin/streptomycin in F-12 medium (Gibco, Waltham, MA, USA). HaCaT are healthy human skin keratinocytes and were cultivated in Dulbecco’s Modified Eagle Medium with a glucose concentration of 4.5 g/L (DMEM (1X) + GlutaMAX − I; Gibco, Waltham, MA, USA) supplemented with 10% FBS and 1% penicillin/streptomycin. All cells were incubated at 37 °C in a humidified 5% CO₂ atmosphere.
CC-115, AZD7648, and Sapanisertib (Selleckchem, Houston, TX, USA) were diluted in dimethyl sulfoxide (DMSO, Roth, Karlsruhe, Germany) and stored at −80 °C. The selected concentrations of KI were determined via a dilution series using CAL33 cells. Required aliquots were freshly thawed prior to each experiment.