Ccd 18co

Human lung carcinoma cell line A549 and CCD-18Co colon fibroblasts were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). The A549 cells were cultured in a complete RPMI-1640 medium (Sigma-Aldrich, St. Louis, MO, USA), and CCD-18Co cells were cultured in an MEM medium (PAN-Biotech GmbH, Aidenbach, Germany). The media were supplemented with 10% fetal bovine serum (FBS; Biosera, Nuaille, France) and antibiotics (1% Antibiotic-Antimycotic 100× and 50 × 10⁻³ g l⁻¹ gentamicin; Biosera) at 37 °C, 95% humidity, and 5% CO₂.
Prior to the selected treatments, cells were seeded on 12-well μ-Chamber slides (ibidi GmbH, Martinsried, Germany) and 6 and/or 96-well plates (TPP, Trasadingen, Switzerland) and left to settle for 24 h. The biscoumarin derivative solutions (at concentrations ranging from 10–100 µM) were then added to cells for 24 or 48 h, and analysis was subsequently performed.

We utilized seven cell lines in this study. The cell lines K562, SK-hep-1, HepG2, and CCD18co were purchased from the American Type Culture Collection (ATCC, Manassas, Virginia, USA). Whereas the cell lines Jurkat, U266 and OPM2 were acquired from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). We cultured K562, U266, Jurkat, and OPM2 in RPMI1640 medium (Pan-Biotech, Aidenbach, Bavaria, Germany) supplemented with 10% FBS (Sigma-Aldrich Chemie GmbH, Munich, Germany) and 1% penicillin/streptomycin (P/S) (Gibco, Schwerte, Germany). While SK-hep-1, HepG2, and CCD18co cells were maintained in EMEM medium (Pan-Biotech, Aidenbach, Bavaria, Germany) supplemented with 10% FBS (Sigma-Aldrich Chemie GmbH, Munich, Germany) and 1% penicillin/streptomycin (P/S) (Gibco, Schwerte, Germany). Meticrane (Sigma-Aldrich Chemie GmbH, Munich, Germany) was dissolved in DMSO and stored at -20°C at a concentration of 200mM. The HDAC inhibitor CUDC-101 (Selleck Chemicals GmbH, Munich, Germany) and the selective HDAC6 inhibitor ACY1215 (Cayman Chemical, Ann Arbor, Michigan, US) was dissolved in DMSO and stored at -20°C at a concentration of 50mM. Also, DNMT1 inhibitor 5-Azacytidine (5AC) (STEMCELL Technologies Germany GmbH, Cologne, Germany) was dissolved in DMSO and stored at -20°C at a concentration of 25mM.

Human lung carcinoma cell line A549 and CCD-18Co colon fibroblasts were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). The A549 cells were cultured in a complete RPMI-1640 medium (Sigma-Aldrich, St. Louis, MO, USA) and CCD-18Co cells were cultured in a minimum essential medium (MEM) (PAN-Biotech GmbH, Aidenbach, Germany) at 37 °C, 95% humidity and 5% CO₂. The media were supplemented with 10% fetal bovine serum (FBS, Biosera, Nuaille, France) and antibiotics (1% Antibiotic-Antimycotic 100 × and 50 × 10⁻³ g L⁻¹ gentamicin, Biosera). Prior to the selected treatments, cells were seeded on 6- and/or 96-well plates (TPP, Trasadingen, Switzerland) and left to settle for 24 h. The acridine compounds solutions (at concentrations ranging from 5–75 µM) were then added to cells for 24 or 48 h, and analysis was subsequently performed.