Mrc 5

The starting materials and other reagents for synthesis were purchased from Aldrich Company and Tokyo Chemical Industry and used as received without further purification unless otherwise indicated. The melting points were measured using an SMP3 StuartTM digital melting point apparatus from Bibby Sterlin, Ltd (Stone, United Kingdom). All synthesized compounds were analyzed using nuclear magnetic resonance (NMR) and mass spectrometry to confirm their structures. Proton and carbon NMR spectra were accomplished using the Bruker Avance III HD 300 spectrometer at 75, 100, 300, and 400 MHz. High-resolution mass spectra were measured with a micrOTOF electrospray ionization time-of-flight mass spectrometer (Bruker Daltonics, Germany). HepG2 (hepatocarcinoma), MOLT-3 (acute lymphoblastic leukemia), HuCCA-1 (cholangiocarcinoma), A549 (lung carcinoma), MRC-5 (normal embryonic lung cell), MDA-MB-231 (hormone-independent breast cancer), S102 (Thai liver cancer), HeLA (cervical carcinoma), T47-D (hormone-dependent breast cancer), H69AR (lung cancer, multidrug resistant), and HL-60 cell lines (promyeloblast) were purchased from Hyclone Laboratories. Doxorubicin (purity ≥98%) and etoposide (purity 98%) were purchased from Aldrich Company.

Colorectal cancer cells HCT116wt and derived cells (HCT116-FUR, HCT116-PROP), and MRC5 fibroblasts were routinely cultured in DMEM culture medium supplemented with 10% FCS (HyClone Laboratories, Logan, UT, USA) and gentamicin (Sandoz, Basel, Switzerland). Experiments in hypoxia (1% O₂) were done in a humidified anaerobic workstation (Ruskinn Technologies, Bridgend, UK) with 5% CO₂, 10% H₂ and 84% N₂ at 37 °C. MRC5 was purchased from ATCC, HCT116 cell line was a gift from Dr. Nicholas Denko.