Fig. 2

RuCUR (ruthenium-curcumin) compound reduced mutant (mut) p53 levels that correlated with reduced proliferation and increased cell death. (a) Mutp53-carrying T98 and SKBR3 cells were seeded on ultra-low attachment multiwell plates allowing for tumor spheroid formation. Four days after seeding, spheroids were formed (approximate size 300–500 μm) and then treated with different doses of RuCUR (10, 50, 100 μM) for the indicated times. Tumor spheroids volume was quantified according to the formula: V = a x (b2)/2, where a and b are, respectively, length and width. Representative images of spheroids derived from both cell lines are shown in the upper panels. Spheroids volumes are reported on the bottom panels. Histograms represent the fold increase quantified with respect to controls set to 1.0, ± SD. * (p ≤ 0.01), # (p ≤ 0.05) (single treatments compared to untreated spheroids). (b) Cell viability was measured by trypan blue exclusion assay in T98 and SKBR3 cells treated for 24 h with different doses of RuCUR (1, 10, 50, 100 μM) and expressed as cell death percentage ± S.D. * (p ≤ 0.01), # (p ≤ 0.05) (single treatments compared to untreated cells). (c) Western blot analysis of p53 protein was performed in T98 and SKBR3 cells untreated or treated with RuCUR (1, 10, 50, 100 μM) for 24 h. The ratio of p53 levels vs β-actin, following densitometric analysis using ImageJ software, is shown. (d) Western blot analysis of the indicated protein levels was performed in T98 and SKBR3 cells untreated or treated with RuCUR (100 μM) for 24 h. Actin was used as protein loading control. The ratio of the protein levels vs β-actin, following densitometric analysis using ImageJ software, is reported. n.s = not specific