Olaparib

Mouse preosteoblastic MC3T3-E1 cells were obtained from RIKEN BRC Cell Bank (Tsukuba, Japan). The cells were maintained in Minimum Essential Medium (no ascorbic acid) (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific, Tokyo, Japan) and 1% penicillin-streptomycin (Nacalai Tesque, Kyoto, Japan). To induce osteoblast differentiation, MC3T3-E1 cells were seeded on 6-, 48-, or 96-well plates at a density of 6.6 × 10³ cells/cm² and cultured for 2 days in maintenance medium. The medium was then changed to differentiation medium containing 1% ascorbic acid, 0.2% hydrocortisone, 2% β-glycerophosphate, 10% FBS (Gibco), and 1% penicillin-streptomycin (Nacalai Tesque) in the presence of olaparib, PDD00017273, or dimethylsulfoxide (DMSO, solvent). The differentiation medium was replaced every 4 days, and the cells were allowed to differentiate for 0, 7, 14, 21, and 28 days for various experiments. The cells were cultured in a humidified atmosphere containing 5% CO₂ at 37 °C. olaparib was purchased from ChemScene (Monmouth Junction, NJ, USA), and PDD00017273 [24 (link)] was purchased from Cayman Chemical Company (Ann Arbor, MI, USA).

U251, U87, and SF767 cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin at 37 °C in a 5% CO₂ atmosphere. TMZ-resistant clones (U251, U87) were derived from the TMZ-sensitive U251 or U87 cells by culturing them with increasing doses of TMZ, as described previously [12 (link)].
Cells were treated with each reagent: TMZ (LKT Laboratories, St. Paul, MN, USA), VAL-083 (MedChemExpress, Shanghai, China), talazoparib, veliparib, MK-8776 (Selleck Chemicals, Houston, TX, USA), and Olaparib (ChemScene, Monmouth Junction, NJ, USA).

The human pancreatic cancer cell line MIA PaCa-2 and C1 cells established in this study were cultured in minimal essential medium (MEM)(Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS) (Cosmobio, Tokyo, Japan), 1% non-essential amino acids (Nacalai Tesque), and 1% penicillin-streptomycin (Nacalai Tesque). Cells were maintained in a humidified atmosphere with 5% CO₂ at 37°C. The PARP inhibitors olaparib and talazoparib were purchased from Chemscene LLC (Monmouth Junction, NJ, USA) and MedChem Express (Monmouth Junction, NJ, USA), respectively. FK866 was obtained from Adipogen Life Sciences, and nicotinamide was purchased from Nacalai Tesuque.