Derivatives of 6-cinnamamido-quinoline-4-carboxamide impair lysosome function and induce apoptosis

// Hsiao-Hui Kuo 1, * , Rajesh Kakadiya 2, * , Yi-Chen Wu 1 , Tsann-Long Su 2 , Te-Chang Lee 2 , Yi-Wen Lin 2 , Ling-Huei Yih 1 1 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan 2 Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan * These authors have contributed equally to this work Correspondence to: Tsann-Long Su, email: tlsu@ibms.sinica.edu.tw Ling-Huei Yih, email: lhyih@gate.sinica.edu.tw Keywords: anticancer, autophagy, cinnamamide quinolines, ERK, lysosome Received: October 23, 2015     Accepted: May 02, 2016     Published: May 13, 2016 ABSTRACT Autophagy is a lysosomal degradative process that protects cancer cells from multiple types of stress. In this study, we synthesized a series of derivatives of 6-cinnamamido-quinoline-4-carboxamide (CiQ), and investigated their effects on the proliferation and autophagy of cancer cells in vitro . These derivatives effectively inhibited the proliferation of a broad spectrum of cancer cell lines. Further study revealed that CiQ derivatives may induce autophagy and result in disruption of autophagy propagation. Consequently, these derivatives triggered massive apoptosis, as evidenced by caspase-9 activation and PARP cleavage. Blockage of autophagy by depletion of autophagy related gene ATG5 or BECN1 considerably alleviated CiQ-induced cell death, indicating that autophagy may mediate CiQ-induced cell death. Furthermore, treatment with CiQ derivatives increased lysosome membrane permeability (LMP) and enhanced accumulation of ubiquitinated proteins, which collectively indicate impaired lysosome function. In addition, treatment of cells with CiQ derivatives activated extracellular signal-regulated kinase (ERK); abrogation of ERK activation, either by treating cells with U0126, an inhibitor of mitogen-activated protein/ERK kinase 1 (MEK1), or by ectopically overexpressing a dominant-negative MEK1, significantly reduced CiQ derivative-induced LMP, LC3 and p62 accumulation, and cytotoxicity. These results indicate that CiQ derivatives activate ERK and disrupt lysosome function, thereby altering autophagic flux and resulting in apoptotic cell death.