N-Benzoyl-O-(N'-(1-benzyloxycarbonyl-4-piperidiylcarbonyl)-D-phenylalanyl)-D-phenylalaninol (
BBP), a novel synthesized
asperphenamate derivative with the increased solubility, showed growth inhibitory effect on human
breast carcinoma MCF-7 cells in a time- and concentration-dependent manner. The growth inhibitory effect of
BBP was associated with induction of autophagy, which was demonstrated by the development of acidic vesicular organelles, cleavage of LC3 and upregulation of Atg4 in
BBP-treated MCF-7 cells. Since the application of Atg4
siRNA totally blocked the cleavage of LC3, we demonstrated a central role of Atg4 in
BBP-induced autophagy. The further studies showed that
BBP increased the levels of
reactive oxygen species (ROS), and pretreatment with NAC effectively blocked the accumulation of ROS, autophagy and growth inhibition triggered by
BBP. Moreover,
BBP induced the activation of JNK, and JNK inhibitor
SP600125 reversed autophagy, the increase of Atg4 levels, conversion of LC3 and growth inhibition induced by
BBP. Knockdown of JNK by
siRNA efficiently inhibited ROS production and autophagy, but
antioxidant NAC failed to block JNK activation induced by
BBP, indicating that JNK activation may be a upstream signaling of ROS and should be a core component in
BBP-induced autophagic signaling pathway. These results suggest that
BBP produces its growth inhibitory effect through induction of the autophagic cell death in MCF-7 cells, which is modulated by a JNK-dependent Atg4 upregulation involving ROS production.