Artesunate, the active agent from Artemisia annua L. used in the
traditional Chinese medicine, is being applied as a first-line
drug for
malaria treatment, and trials are ongoing that include this
drug in
cancer therapy. Despite increasing interest in its therapeutic application, the mode of cell killing provoked by
artesunate in human cells is unknown. Here, we show that
artesunate is a powerful inducer of oxidative DNA damage, giving rise to
formamidopyrimidine DNA glycosylase-sensitive sites and the formation of
8-oxoguanine and 1,N6-ethenoadenine. Oxidative DNA damage was induced in LN-229 human
glioblastoma cells dose dependently and was paralleled by cell death executed by apoptosis and
necrosis, which could be attenuated by radical scavengers such as N-acetyl
cysteine. Oxidative DNA damage resulted in
DNA double-strand breaks (
DSB) as determined by γH2AX foci that colocalized with 53BP1. Upon chronic treatment with
artesunate, the level of
DSB continuously increased over the treatment period up to a steady-state level, which is in contrast to ionizing radiation that induced a burst of
DSB followed by a decline due to their repair. Knockdown of Rad51 by
short interfering RNA and inactivation of
DNA-PK strongly sensitized
glioma cells to
artesunate. These data indicate that both homologous recombination and nonhomologous end joining are involved in the repair of
artesunate-induced
DSB.
Artesunate provoked
a DNA damage response (DDR) with phosphorylation of ATM, ATR, Chk1, and Chk2. Overall, these data revealed that
artesunate induces oxidative DNA lesions and
DSB that continuously increase during the treatment period and accumulate until they trigger DDR and finally
tumor cell death.