Investigating Osteosarcoma and Malignant Mesothelioma Cells

U-2OS human osteosarcoma cells were purchased from the American Type Culture Collection (ATCC) and grown in IMDM supplemented with 10 % FBS, 2 mM l-glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin (EuroClone, Milan, Italy). Experiments were also performed with MM-B1 biphasic malignant mesothelioma cells [18 (link)] and with the HT-29 and COLO 205 colorectal adenocarcinoma and MCF-7 breast adenocarcinoma cells of the NCI-60 cell line panel. MCF-7, COLO 205 and HT-29 cells were grown in RPMI 1640, and MM-B1 in DMEM (EuroClone), both media being supplemented as above. NBDHEX and MC3181 were synthesized as previously reported [1 (link), 5 (link)]. For cell treatments NBDs were dissolved in DMSO and diluted in cell medium, with the final DMSO concentration never exceeding 0.01 % (v/v); control cultures received an equivalent amount of DMSO vehicle. The late-stage autophagy inhibitor chloroquine diphosphate (CQ) was purchased from Sigma-Aldrich (Milan, Italy).