Reversible anti-proliferative activity of bDHC on human normal cells.

<p><b>A.</b> PI/FACS analysis of HF cell cycle progression after DMSO and bDHC treatments for 24 and 48 hours (left and middle panel, respectively). PI/monoparametric cell cycle analysis of bDHC-treated cells <i>versus</i> bDHC-released cells (right panel) (A = SubG1, B = G0/G1, C = S, D = G2/M). <b>B.</b> Left and middle panels: Analysis of cell cycle progression of WRL68 cells following 24 and 48 hours of treatment with DMSO or bDHC. Right panel: PI/FACS monoparametric analysis of WRL68 cells treated with bDHC for 48 hours and then released into fresh medium for additional 24 hours (A = SubG1, B = G0/G1, C = S, D = G2/M). <b>C.</b> Quantitative evaluation of bDHC concentration by UV-vis spectroscopy. Upper left panel: bDHC recovered from cell lysates of HCT116 (•) and HF (▴) cells after 24 hours treatment at different concentrations (10, 20 and 30 µM). bDHC recovered from cell pellets (ng) was referred to total cellular proteins (mg), determined using Bradford method. Upper right panel: Comparison of cellular uptake (20 and 30 µM concentrations) in HCT116 (black histograms) and HF cells (grey histograms). Lower panel: bDHC recovered from culture media following incubation of HCT116 (•) and HF (▴) cells with bDHC at 10, 20 and 30 µM concentration. All data were normalized on control (DMSO). Drug amount was determined by reading absorbance at λ_max = 393 nm. Reported values are an average of three independent experiments −/+ SD.