Although
eupafolin, a
flavone found in Artemisia princeps Pampanini, has been shown to inhibit the growth of several human
cancer cells, its mode of action is poorly understood. In this study, we investigated the pro-apoptotic activities of
eupafolin in human cervical
carcinoma HeLa cells. It was found that
eupafolin induced apoptosis in a dose-dependent manner, as evidenced by DNA fragmentation and the accumulation of positive cells for
annexin V. In addition,
eupafolin triggered the activations of caspases-3, -6, -7, -8, and -9 and the cleavages of their substrates, such as,
poly (ADP-ribose) polymerase and
lamin A/C. Furthermore, treatment with
eupafolin resulted in a loss of mitochondrial membrane potential (DeltaPsi(m)), increased the release of
cytochrome c to the cytosol, and altered the expression levels of
B-cell lymphoma 2 (Bcl-2) family
proteins. Interestingly,
caspase-8, an initiator
caspase, was activated after the loss of DeltaPsi(m) and the activations of caspases-3 and -9. Moreover, treatment with
z-DEVD-fmk (a specific
caspase-3 inhibitor) and the overexpression of Bcl-2 prevented
eupafolin-stimulated
caspase-8 activation. Altogether, these results suggest that the
eupafolin-induced apoptosis in HeLa cells is mediated by
caspase-dependent pathways, involving caspases-3, -9, and -8, which are initiated by the Bcl-2-dependent loss of DeltaPsi(m).