Augmentation of Curcumin Induced Cell Death by Cell Permeable C2 Ceramide in Human T-lymphoma Jurkat Cell Line

Document Type : Original Article

Author

Laboratory of Molecular Cell Biology, Department of Zoology, Faculty of Science, Assiut University, Assiut 71516, Egypt

Abstract

Lymphoma in general is hard to treat cancer type which is resistant to be cured with variety of anticancer agents. Consequently, finding new approaches for anti-lymphoma agents is urgent. The present study evaluates the augmentation effect of cell permeable short chain C2 ceramide in sensitization of Human T-lymphoma Jurkat cells to curcumin-induced cell death. Many methods were used in the present study include MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide) assay for determination of cell viability, DAPI (4′,6-diamidino-2-phenylindole ) and SRB (sulforhodamine B) methods for determination of cell death, Immunofluorescence staining of endogenous ceramide and western blot detection of apoptosis associated proteins. Curcumin alone reduces cell viability and induces apoptotic cell death in Jurkat cell line as indicated from cleavage of the proapoptotic caspase3 and cleavage of [poly (ADP-ribose) polymerase] PARP. C2 ceramide potentiates the apoptotic cell death induced by curcumin. Hence the combination of curcumin and C2 ceramide dramatically increases the apoptotic cell death associated proteins; cleaved caspase 3 and PARP in addition to apoptotic index obtained from DAPI and SRB assays. Moreover, the molecular mechanism of C2 ceramide sensitization of apoptotic cell death induced by curcumin was outlined. 

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