Reactive oxygen species (ROS) are produced by all aerobic cells and have been implicated in the regulation of diverse cellular functions, including intracellular signaling, transcription activation, proliferation, and apoptosis.
Salvicine, a novel
diterpenoid quinone compound, demonstrates a broad spectrum of antitumor activities. Although
salvicine is known to trap the
DNA-topoisomerase II (
Topo II) complex and induce
DNA double-strand breaks (DSBs), its precise antitumor mechanisms remain to be clarified. In this study, we investigated whether
salvicine altered the levels of ROS in
breast cancer MCF-7 cells and whether these ROS contributed to the observed antitumoral activity. Our data revealed that
salvicine stimulated intracellular ROS production and subsequently elicited notable DSBs. The addition of N-acetyl
cysteine (NAC), an
antioxidant, effectively attenuated the
salvicine-induced ROS enhancement and subsequent
DNA DSBs. Heat treatment reversed the accumulation of
DNA DSBs, and the addition of NAC attenuated the
Topo II-
DNA cleavable complexes formation and the growth inhibition of
salvicine-treated JN394top2-4 yeast cells, collectively indicating that
Topo II is a target of the
salvicine-induced ROS. On the other hand, when examining the impact of
salvicine on DNA repair pathways, we unexpectedly observed that
salvicine selectively down-regulated the catalytic subunit of
DNA-dependent protein kinase (
DNA-PK(CS))
protein levels and repressed
DNA-PK
kinase activity; both of these effects were attenuated by NAC pretreatment of MCF-7 cells. Finally and most importantly, NAC attenuated
salvicine-induced apoptosis and cytotoxicity in MCF-7 cells. These results indicate that apart from its direct actions,
salvicine generates ROS that modulate DNA damage and repair, contributing to the comprehensive
biological consequences of
salvicine treatment, such as
DNA DSBs, apoptosis, and cytotoxicity in
tumor cells. The finding of
salvicine-induced ROS provides new evidence for the molecular mechanisms of this compound. Moreover, the effects of
salvicine-induced ROS on
Topo II and
DNA-PK give new insights into the diverse
biological activities of ROS.