Figure 2. Ac-LA induces apoptosis in prostate cancer cells.

(A) Ac-LA induced translocation of phosphatidylserine to the outer leaflet of the cell membrane. PC-3 cells were treated for 24 h, stained with annexin V-FITC (green) and PI (red), and analyzed by confocal fluorescence microscopy. (B) PC-3 cells were treated for different time periods with 10 μM ac-LA, stained as in A and analyzed by FACS. (C) Ac-LA induces DNA laddering in PC-3 cells treated for 48 h with 10 μM ac-LA; 100 μM lupeol and 10 nM docetaxel were used for comparison. (D) Ac-LA increases the apoptotic sub-G₁ cell population. Left panel: typical histograms are shown (treatment for 72 h). (E) Ac-LA induces apoptosis in prostate PC-3 cancer cells more efficiently than in the breast cancer cell line MDA-MB-231. Cell were analyzed as in D after treatment with 10 μM of either ac-LA or lupeol for 72 h. (F) Cell-cycle analysis of cells treated with ac-LA or lupeol as in E. (G) Caspase 3 activation in PC-3 cells treated as in E for 6 h and analyzed using the caspase 3 substrate Z-DEVD-R110 followed by flow cytometry. (H) AcoA induces loss of mitochondrial membrane potential in PC-3 but not MDA-MB-231 cells. Cell were treated as in E for 6 h and the mitochondrial membrane potential, ΔΨm (red/green fluorescence intensity ratio), was analyzed using JC-1 dye staining followed by flow cytometry. All results are mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, n = 3.