A2780 cell line

The human ovarian carcinoma A2780 cell line was purchased from ATCC. A2780 sublines that were resistant to CIS (A2780CR1, A2780CR2 (A2780 cisplatin-resistant)), and PAC (A2780PR1 and A2780PR2 (A2780 paclitaxel-resistant)) were obtained by exposing A2780 cells to the relevant drugs at gradually increasing concentrations. The final concentrations used to select the resistant cells were 1000 ng/mL of CIS for both A2780CR1 and A2780CR2 cell lines, 300 ng/mL of PAC for A2780PR1 cell line, and 1100 ng/mL of PAC for A2780PR2 cell line, as described previously [39 (link)]. They were twofold greater than the concentrations in the plasma 2 h after intravenous administration [92 (link)]. According to parental drug-sensitive cell lines, the increase in resistance was as follows: 4.09-fold for A2780CR1 vs. A2780, 3.29-fold for A2780CR2 vs. A2780, 146-fold for A2780PR1 vs. A2780, and 1202-fold for A2780PR2 vs. A2780, as we described previously [39 (link)]. All cell lines were maintained as a monolayer in the complete medium (MEM medium supplemented with 10% (v/v) fetal bovine serum, 2 pM L-glutamine, penicillin (100 U/mL), streptomycin (100 U/mL) and amphotericin B (25 µg/mL)) at 37 °C in a 5% CO₂ atmosphere.

Vero cell (derived from the kidney of African green monkey), PC-3 cell line (Prostate carcinoma), and A2780 cell line (Ovarian carcinoma) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) boosted with 10% heat-inactivated fetal bovine serum, 1% l-glutamine, buffer of HEPES and 50 µg/mL gentamycin. The cells were maintained at 37 °C in a humidified 5% CO₂ atmosphere and were subcultured twice a week [26 (link)].

The A2780 cell line was purchased from American Type Culture Collection (ATCC), and SAHA was purchased from Selleckchem (Houston, TX, USA). Primary antibodies against Ac-H2A, Ac-H2B, Ac-H3, Ac-H4, HDAC2, HDAC3, HDAC4, DNMT3A, PRMT1, SUV39H1, MDMX, p53, p21^WAF1/CIP1, p27^Kip1, AURKB, CDC25C, GADD45A (1:1000) and Alexa Fluor 488 dye were purchased from Cell Signaling Technology (Danvers, MA, USA). The MDM2 antibody (1:500) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). β-actin (1:5000), secondary antibodies (anti-rabbit, anti-mouse), horseradish peroxidase (HRP) conjugate, and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich (St. Louis, MO, USA). Nitrocellulose membrane (NC) (0.45 µm) was purchased from Amersham (GE Healthcare Life Sciences, Marlborough, MA, USA) and KPL LumiGlo Reserve chemiluminescent substrate was purchased from SeraCare Life Sciences (Milford, MA, USA).

The human ovarian cancer A2780 cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA). The cells were grown in RPMI1640 medium (Cellgro, Manassas, VA) supplemented with 10% fetal bovine serum (Gibco BRL, Carlsbad, MD), 100 units/mL penicillin, and 100 μg/mL streptomycin. They were incubated at 37°C in a humidified atmosphere with 5% CO₂. The Cell Counting Kit-8 (CCK-8, Dojindo Laboratories, Japan) was used to determine cell proliferation according to the manufacturer's recommendations. Briefly, cells were seeded in 96-well plates at a density of 1 × 10⁴ cells/well and incubated for 24 h at 37°C. The cells were treated with different concentrations of compounds. After incubation for 24 h, 10 μL of the kit reagent was added to each well, and the cells were incubated for an additional hour. Cell proliferation was measured by scanning with a microplate reader at 450 nm. Control cells were exposed to culture media containing 0.5% v/v dimethyl sulfoxide (DMSO).