JNK-dependent Atg4 upregulation mediates asperphenamate derivative BBP-induced autophagy in MCF-7 cells

• Published in: Toxicology and applied pharmacology 2012-08, Vol.263 (1), p.21-31
• Main Authors: Li, Yanchun, Luo, Qiyu, Yuan, Lei, Miao, Caixia, Mu, Xiaoshuo, Xiao, Wei, Li, Jianchun, Sun, Tiemin, Ma, Enlong
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Antineoplastic Agents - pharmacology; ANTIOXIDANTS; APOPTOSIS; Apoptosis - drug effects; Asperphenamate; Atg4; Autophagy; Autophagy - drug effects; Autophagy-Related Proteins; Benzamides - pharmacology; Biological and medical sciences; Blotting, Western; Breast Neoplasms - drug therapy; Carcinoma - drug therapy; CARCINOMAS; CELL CONSTITUENTS; Cell Cycle - drug effects; Cell Line, Tumor; CONCENTRATION RATIO; Cysteine Endopeptidases - physiology; Dose-Response Relationship, Drug; Female; HeLa Cells; Humans; INHIBITION; JNK; JNK Mitogen-Activated Protein Kinases - physiology; MAMMARY GLANDS; Medical sciences; Phenylalanine - analogs & derivatives; Phenylalanine - pharmacology; Reactive oxygen species; Reactive Oxygen Species - metabolism; Toxicology; TUMOR CELLS; Up-Regulation - drug effects
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1016/j.taap.2012.05.018

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. ► Asperphenamate derivative BBP with increased solubility was synthesized. ► BBP selectively inhibited the growth of human breast tumor cells. ► The growth inhibitory effect of BBP was associated with induction of autophagy. ► JNK-dependent Atg4 upregulation mediated BBP-induced autophagy.