Cell Cycle Analysis of MCF-7 Cells

The effect of compounds 9 and 13 and doxorubicin (positive control) in cell cycle distribution of MCF-7 cells was performed by flow cytometry using BD cycletest™ Plus DNA Kit Assay (BD Biosciences, San Jose, USA) as previously described [23 (link)]. Cells were tested in 6-well plates (3 mL, 1 × 10⁵ cells/mL) and the incubation time was 72 h in humidified 5% CO₂ atmosphere at 37 °C. The tested concentrations were ¼ × IC₅₀, ½ × IC₅₀ and IC₅₀. Untreated cells (control) were used for comparison with treated cells. The BD FACS Aria I Cell Sorter Flow Cytometer (Becton-Dickinson, Germany) was then used for cell cycle analysis. For each sample, 10⁴ cells were counted. For PI excitation, an argon-ion laser emitting at 488 nm was used. Cytographs were analyzed using BD FACSDiva™ Flow Cytometry Software Version 6.1.2 (Becton-Dickinson).