Figure 5

ML-9 induces prostate cancer cell death. (a) ML-9 reduced cell viability in a concentration-dependent manner. LNCaP cells, plated on 96-well plate, were treated with different doses of ML-9 in full or serum-starved media for 24 h. Cell viability was monitored using the CellTiter 96 Aqueous One Solution cell proliferation assay (MTS assay). Following treatment, the cells were incubated with reagent solution and absorbance was recorded at 490 nm wavelength using an ELISA plate reader. Values represent means±S.E.M. n=3. (b) Cell viability was assessed by trypan blue exclusion assay. LNCaP cells were treated with different concentrations of ML-9 in full or serum-starved media for 24 h, the cells were collected by trypsin-EDTA, incubated with trypan blue and viable cells as well as dead (blue) cells were counted using haemocytometer. Values represent means±S.E.M. ⁺⁺⁺P<0.001. n=4. (c) ML-9 induces PARP cleavage. LNCaP cells were treated with full, serum-starved, 30 μM ML-9-containing full medium or 30 μM ML-9-containing serum-starved medium for 24 h. (d) ML-9 induces apoptosis in LNCaP cells. LNCaP cells were treated as in (c). At the end of the treatment, cells were collected, stained with Annexin V/PI and subjected to fluorescence microscopy analysis. Data are represented as means±S.E.M. n=4. (e) LNCaP, PC3 and DU145 cells, plated on 96-well plate, were treated with docetaxel (5 nM), ML-9 (30 μM) or combination of docetaxel+ML-9 for 48 h. Cell viability was monitored using the CellTiter 96 Aqueous One Solution cell proliferation assay (MTS assay)