Novel derivatives of spirohydantoin induce growth inhibition followed by apoptosis in leukemia cells

Kavitha, CV and Nambiar, Mridula and Kumar, Ananda CS and Choudhary, Bibha and Muniyappa, K and Rangappa, Kanchugarakoppal S. and Raghavan, Sathees C (2009) Novel derivatives of spirohydantoin induce growth inhibition followed by apoptosis in leukemia cells. In: Biochemical Pharmacology, 77 (3). pp. 348-363.

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Abstract

Hydantoin derivatives possess a variety of biochemical and pharmacological properties and consequently are used to treat many human diseases. However, there are only few studies focusing on their potential as cancer therapeutic agents. In the present study, we have examined anticancer properties of two novel spirohydantoin compounds, 8-(3,4-difluorobenzyl)-1'-(pent-4-enyl)-8-azaspiro[bicyclo[3.2.1] octane-3,4'-imidazolidine]-2',5'-dione (DFH) and 8-(3,4-dichlorobenzyl)-1'-(pent-4-enyl)-8-azaspiro[bicyclo[3.2.1]octane-3,4'-imidazolidine]-2',5'-dione (DCH). Both the compounds exhibited dose- and time-dependent cytotoxic effect on human leukemic cell lines, K562, Reh, CEM and 8ES. Incorporation of tritiated thymidine ([H-3) thymidine) in conjunction with cell cycle analysis suggested that DFH and DCH inhibited the growth of leukemic cells. Downregulation of PCNA and p-histone H3 further confirm that the growth inhibition could be at the level of DNA replication. Flow cytometric analysis indicated the accumulation of cells at subG1 phase suggesting induction of apoptosis, which was further confirmed and quantified both by fluorescence-activated cell sorting (FACS) and confocal microscopy following annexin V-FITC/propidium iodide (PI) staining. Mechanistically, our data support the induction of apoptosis by activation of the mitochondrial pathway. Results supporting such a model include, elevated levels of p53, and BAD, decreased level of BCL2, activation and cleavage of caspase 9, activation of procaspase 3, poly (ADP-ribosyl) polymerase (PARP) cleavage, downregulation of Ku70, Ku80 and DNA fragmentation. Based on these results we discuss the mechanism of apoptosis induced by DFH and its implications in leukemia therapy. (C) 2008 Elsevier Inc. All rights reserved.

Item Type:	Journal Article
Publication:	Biochemical Pharmacology
Publisher:	Elsevier
Additional Information:	Copyright of this article belongs to ElsevierScience.
Keywords:	Chemotherapy;Double-strand break;Cytotoxicity;Cell death;Anticancer drugs;Flow cytometry;DNA damage.
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	05 Nov 2009 09:48
Last Modified:	19 Sep 2010 05:27
URI:	http://eprints.iisc.ac.in/id/eprint/19206

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