Novel Rhodanine Derivative 5-4-(4-Fluorophenoxy) phenylmethylene-3-{4-3-(4-methylpiperazin-1-yl) propoxyphenyl}-2-thioxo-4-thiazolidinone dihydrochloride Induces Apoptosis via Mitochondria Dysfunction and Endoplasmic Reticulum Stress in Human Colon Cancer Cells

摘要

We previously reported that 5-[4-(4-fluorophenoxy) phenyl] methylene-3-{4-[3-(4-methylpiperazin-1-yl)propoxy]phenyl}-2-thioxo-4-thiazolidinone dihydrochloride () has potent, selective and metabolically stable IKKβ inhibitory activities. However, the apoptosis-inducing of and its underlying mechanism have not yet been elucidated in human colon cancer cells. We show that triggered apoptosis, as indicated by externalization of Annexin V-targeted phosphatidylserine residues in HT-29 and HCT-116 cells. induced the activation of caspase-8, -9, and -3, and the cleavage of poly (ADP ribose) polymerase-1 (PARP-1). -induced apoptosis was significantly suppressed by pretreatment with z-VAD-fmk (a broad caspase inhibitor). also induced release of cytochrome c (Cyt c), apoptosis inducing factor (AIF), and endonuclease G (Endo G) by damaging mitochondria, resulting in caspase-dependent and -independent apoptotic cell death. triggered endoplasmic reticulum (ER) stress and changed the intracellular calcium level ([Ca²⁺]i). Interestingly, treatment with activated not only ER stress marker proteins including inositol-requiring enzyme 1-alpha (IRE-1α) and protein kinase RNA-like endoplasmic reticulum kinase (PERK), but also μ-calpain, and caspase-12 in a time-dependent manner. -induced apoptosis substantially decreased in the presence of BAPTA/AM (an intracellular calcium chelator). Taken together, these results suggest that mitochondrial dysfunction and ER stress contribute to -induced apoptosis in human colon cancer cells.