Skov 3 human ovarian cancer cells

All the cells used in the present work were SKOV-3 human ovarian cancer cells (ATCC, Manassas, U.S.A.). The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic solution (Gibco, Invitrogen, US) at 37 °C in a humidified 5% CO₂ incubator. Before fluorescence imaging, the cells were isolated at the exponential growth phase with trypsin solution (1% poly-vinyl pyrrolidone, 0.2% EGTA and 0.25% trypsin containing 0.02% EDTA) at 37 °C for three minutes. After washing with phosphate buffer saline (PBS), the SKOV-3 cells were seeded in 35 mm glass Petri dishes with 3 × 10⁴ cells for the subsequent experiments.

SK-OV3 human ovarian cancer cells were purchased from American Type Culture Collection. The cells were cultured in complete RPMI-1640 medium (Thermo Fisher Scientific Inc.) containing 10% fetal bovine serum (Gibco; Thermo Fisher Scientific Inc.) and no antibiotics at 37°C and 5% humidified CO₂. Cells in the logarithmic growth phase were used for the experiments. Thiazolyl blue tetrazolium bromide (MTT) was obtained from Abcam (cat. no. ab211091). The primary antibodies GADD45α (cat. no. ab203090), Ser-139 phosphorylated H2AX (γ-H2AX; cat. no. ab26350), H2AX (cat. no. ab229914) and β-actin (cat. no. ab8226), all 1:1,000, were purchased from Abcam. CDDP was acquired from Sigma-Aldrich (Merck KGaA).

All chemical spectral analyses were conducted at Kuwait University Research Center. Thin layer chromatography (TLC) was performed using Polygram SIL G/UV 254 TLC plates, and the results were visualized under ultraviolet light at 254 and 350 nm, with hexane/ethyl acetate as a solvent. Column chromatography was performed using silica gel 60A with a mesh size of 40–60 µm. ¹H and ¹³C nuclear magnetic resonance (NMR) spectra were obtained using a Bruker DPX 600 NMR spectrophotometer at 600 MHz and 150 MHz in DMSO and CDCL3, respectively. Mass spectra were detected on a GC-MS DFS–Thermo spectrometer. An IR spectrum was obtained with a Jasco 6300 FTIR spectrometer. Melting point was determined using a Netzsch DSC 204 F1 Phoenix differential scanning calorimeter.
SKOV3 human ovarian cancer cells were obtained from American Type Culture Collection. A cholesterol-conjugated HSP27 inhibitor was synthesized in our laboratory. Cell culture media and other supplements, such as 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), were obtained from Sigma-Aldrich (Milwaukee, WI). Chemicals and reagents are commercially available and ready for direct use, requiring no preparation. Biological parameters were examined at the Biology Department, Kuwait University.