Calu 1

A549 (NCI), SK-MES-1 and Calu-1 (both ATCC) were cultured in RMPI-1640, supplemented with 10% FBS, 1%Pen/Strep and 1%l-Glutamine (all from Sigma). NIC-H23 cells were cultured in RMPI-1640, supplemented with 10% FBS, 1%Pen/Strep, 1%l-Glutamine and 1 mM sodium pyruvate (Sigma). LL/2, Ladi 3.1 and Ladi 2.1 cells were cultured in DMEM (Sigma) supplemented with 10% FBS, 1%Pen/Strep and 1%l-Glutamine. Human cells were STR-profiled, used between passages 3 and 15, examined for mycoplasma and maintained in Plasmocin (Invivogen) to prevent contamination.

The murine cancer cell lines CT26 (colon) and 4T1 (breast) and the human cancer cell lines Calu-1 (lung), MDA-MB231 (breast), LNCaP (prostate) and IMR-32 (neuroblastoma) were purchased from ATCC (LGC Standards Srl, Milan, Italy). The human melanoma cell line LB24Dagi (LB24) was kindly provided by Dr. D. Castiglia (Istituto Dermopatico dell’Immacolata, Department of Molecular and Cellular Biology, Rome). All cancer cell lines were cultured at 37 ^oC under 5% CO₂ in DMEM medium (Euroclone, Milan, Italy) supplemented with 1% L-glutamine, penicillin/streptomycin, nonessential amino acids and 10% fetal bovine serum (all from Sigma Aldrich). In all cases experiments were performed in triplicate, glucose concentration of administered medium was set at 11.1 mM. MTF treatments were performed for 24 h at concentrations ranging from 1 mM to 10 mM, while CBX was administered at 180 μM for CT26 and at 135 μM for 4T1 on the basis of preliminary assessment of IC50 dose.

Human LUSC H520 and Calu-1 cell lines were originally from ATCC (Manassas, VA, USA). Cells were grown in Roswell Park Memorial Institute (RPMI)-1640 media supplemented with 10% FBS (GIBCO, Invitrogen, USA) and in a humidified atmosphere under 5% CO₂ and 37 °C temperature in a CO₂ incubator (Thermo Fisher Scientific, Waltham, MA, USA).

The human NSCLC cell lines—A549 and Calu-1, breast cancer cell line—T47D, and cervical carcinoma cell line—HeLa, were purchased from ATCC (Rockville, MD). Normal human bronchial epithelial cells—Beas-2B, were kindly provided by Dr M. Rusin from the Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, Poland. A549, Calu-1 and T47D cells were routinely maintained in RPMI 1640 medium, Beas-2B cell line was cultured in DMEM/F12 medium, and HeLa cell line was maintained in DMEM. Cells were grown at 37 °C in humidified air with 5 % CO₂. All culture media were supplemented with 10 % heat-inactivated FBS, 2 mM glutamine and penicillin–streptomycin–amphotericin B solution (10,000 U penicillin, 10 mg streptomycin and 25 μg amphotericin B/ml).

A549 (ATCC® CCL-185™), A427 (ATCC® HTB-53™), and Calu-1 (ATCC® HTB-54™) cell lines were obtained from the ATCC (Manassas, VA, USA), INER-51 (CVCL_5531) was a kind gift from the National Institute of Respiratory Diseases (Mexico). The cells were incubated in a humidified atmosphere with 5% CO₂ at 37 °C and were maintained in culture medium DMEM/F-12 containing 2.50 mM L-Glutamine, 15 mM HEPES buffer medium (Gibco, Grand Island NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco, Grand Island NY, USA) and 100 U/mL penicillin/streptomycin (Gibco, Grand Island NY, USA). The cell lines were grown in plastic tissue-culture dishes (Corning, NY, USA).

Human NSCLC cells (A549, H1975, H292, H522, HCC827, Calu-1, H520, H1299, H460, H1993, H2122, and H1703) were purchased from ATCC (Manassas, VA, USA). Other NSCLC cells were kindly provided by Dr. John V. Heymach (MD Anderson Cancer Center, Houston, TX, USA). PC-9R cells (gefitinib-resistant PC-9 cells) were kindly provided by Dr. Mien-Chie Hung (MD Anderson Cancer Center, Houston, TX, USA). PC-9/ER (H) cells (erlotinib-resistant PC-9 cells) were kindly provided by Dr. Jae Cheol Lee (Asan Medical Center, Seoul, Republic of Korea).The cells were maintained in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and antibiotics. The cells were incubated at 37 °C with 5 % CO₂ in a humidified atmosphere.

All cell lines were of human origin. Melanoma cell lines DOR, Beu, and Hbl were previously described [29 (link)]. Other melanoma cell lines (MeWo, SK-MEL-2, SK-MEL-28, SK-MEL-5, SK-MEL-3, Malme 3M, HT144, WM35, WM1552C, and RPMI-7931) were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA). Normal human melanocytes HeMN-LP were from Cascade Biologics (Portland, OR, USA). NSCLC lung cancer cell lines A549, HT1299, A-427, Calu-1, H-460, H-520, H596, H-661, H-2170, and SK-MES-1, and SCLC cell lines H-446, H-69, H-209, H-82, H-345, H-146, H-378, H-196 were purchased from ATCC. 293FT cells were from Invitrogen (Carlsbad, CA, USA). Colorectal cell lines LoVo, SW480, HCT116 were from ATCC. All other cell lines were purchased also from ATCC: G-401 and A-204 (rhabdoid tumors), U-2 OS and Saos-2 (osteosarcomas), HeLa S3 and C33A (cervical carcinomas), 293 (renal carcinoma), HT-1080 (connective tissue fibrosarcoma), SW-13 (adrenal gland carcinoma), T98G (glioblastoma), IMR90 and WI-38 (normal human fibroblasts), Jurkat (T-cell leukemia), Hep-G2 (hepatocellular carcinoma), SK-N-SH and SH-N-MC (neuroblastomas), PANC-1, PA-TU-8902, MIA PaCa-2, and BxPC-3 (pancreatic carcinomas).