title: Glutathione regulates caspase-dependent ceramide production and curcumin-induced apoptosis in human leukemic cells.
abstract: Depletion of intracellular glutathione (GSH) is the prime hallmark of the progression of apoptosis. Previously, we reported that curcumin induces reactive oxygen species (ROS)-mediated depletion of GSH, which leads to caspase-dependent and independent apoptosis in mouse fibroblast cells (F. Thayyullathil et al., Free Radic. Biol. Med.45, 1403-1412, 2008). In this study, we investigated the antileukemic potential of curcumin in vitro, and we further examined the molecular mechanisms of curcumin-induced apoptosis in human leukemic cells. Curcumin suppresses the growth of human leukemic cells via ROS-independent GSH depletion, which leads to caspase activation, inhibition of sphingomyelin synthase (SMS) activity, and induction of ceramide (Cer) generation. Pretreatment of leukemic cells with carbobenzoxy-Val-Ala-Asp fluoromethylketone, a universal inhibitor of caspases, abrogates the SMS inhibition and Cer generation, and in turn prevents curcumin-induced cell death. Curcumin treatment of leukemic cells also downregulates the expression of the inhibitor of apoptosis proteins (IAPs), phospho-Akt, c-Myc, and cyclin D1. Extracellular supplementation with GSH attenuates curcumin-induced depletion of GSH, caspase-dependent inhibition of SMS, Cer generation, and downregulation of IAPs, whereas, L-D-buthionine sulfoximine, a widely used inhibitor of GSH synthesis, potentiates GSH depletion, Cer generation, and apoptosis induced by curcumin. Taken together, our findings provide evidence suggesting for the first time that GSH regulates caspase-dependent inhibition of SMS activity, Cer generation, and apoptosis induced by curcumin in human leukemic cells.
authors.affiliation: Cell Signaling Laboratory, Department of Biochemistry, Faculty of Medicine and Health Sciences, UAE University, Al Ain, United Arab Emirates.
authors.forename: Jaleel
authors.initials: J
authors.lastname: Kizhakkayil
authors.forename: Faisal
authors.initials: F
authors.lastname: Thayyullathil
authors.forename: Shahanas
authors.initials: S
authors.lastname: Chathoth
authors.forename: Abdulkader
authors.initials: A
authors.lastname: Hago
authors.forename: Mahendra
authors.initials: M
authors.lastname: Patel
authors.forename: Sehamuddin
authors.initials: S
authors.lastname: Galadari
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D020169:Caspases / Q000378:metabolism*; D002460:Cell Line; D002518:Ceramides / Q000096:biosynthesis*; D003474:Curcumin / Q000494:pharmacology*; D005978:Glutathione / Q000502:physiology*; D006801:Humans; D007938:Leukemia / Q000201:enzymology / Q000473:pathology*
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D002518:Ceramides; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D020169:Caspases; D005978:Glutathione; D003474:Curcumin