title: The effect of Amaryllidaceae alkaloids haemanthamine and haemanthidine on cell cycle progression and apoptosis in p53-negative human leukemic Jurkat cells.
abstract: Plants from the Amaryllidaceae family have been shown to be a promising source of biologically active natural compounds of which some selected are currently in pre-clinical development. Regardless of interesting pioneer works, little is known about Amaryllidaceae alkaloids that have shown promising anti-cancer activities. The crinane group of the Amaryllidaceae, including haemanthamine and haemanthidine, was amongst the first of these compounds to exhibit an interesting cytotoxic potential against cancer cell lines. However, the mechanism of cytotoxic and anti-proliferative activity is not yet entirely clear. The primary objectives of the current study were to investigate the effects of haemanthamine and haemanthidine on the induction of apoptosis and the cell cycle regulatory pathway in p53-null Jurkat cells. Results indicate that haemanthamine and haemanthidine treatment decreases cell viability and mitochondrial membrane potential, leads to a decline in the percentage of cells in the S phase of the cell cycle, induces apoptosis detected by Annexin V staining and increases caspase activity. Dose dependent apoptosis was cross verified by fluorescence and bright field microscopy through Annexin V/propidium iodine staining and morphological changes which characteristically attend programmed cell death. The apoptotic effect of haemanthamine and haemanthidine on leukemia cells is more pronounced than that of gamma radiation. Contrary to gamma radiation, Jurkat cells do not completely halt the cell cycle 24h upon haemanthamine and haemanthidine exposure. Both Amaryllidaceae alkaloids accumulate cells preferentially at G1 and G2 stages of the cell cycle with increased p16 expression and Chk1 Ser345 phosphorylation. Concerning the pro-apoptotic effect, haemanthidine was more active than haemanthamine in the Jurkat leukemia cell line.
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, Pardubice 532 10, Czech Republic. Electronic address: radim.havelek@upce.cz.
authors.forename: Radim
authors.initials: R
authors.lastname: Havelek
authors.affiliation: Department of Medical Biochemistry, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Simkova 870, Hradec Kralove 500 38, Czech Republic.
authors.forename: Martina
authors.initials: M
authors.lastname: Seifrtova
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, Pardubice 532 10, Czech Republic.
authors.forename: Karel
authors.initials: K
authors.lastname: Kralovec
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, Pardubice 532 10, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Bruckova
authors.affiliation: ADINACO Research Group, Department of Pharmaceutical Botany and Ecology, Faculty of Pharmacy, Charles University in Prague, Heyrovskeho 1203, Hradec Kralove 500 05, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Cahlikova
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, Pardubice 532 10, Czech Republic.
authors.forename: Marketa
authors.initials: M
authors.lastname: Dalecka
authors.affiliation: Department of Radiobiology, Faculty of Military Health Sciences, University of Defense Brno, Trebesska 1575, Hradec Kralove 500 01, Czech Republic.
authors.forename: Jirina
authors.initials: J
authors.lastname: Vavrova
authors.affiliation: Department of Medical Biochemistry, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Simkova 870, Hradec Kralove 500 38, Czech Republic.
authors.forename: Martina
authors.initials: M
authors.lastname: Rezacova
authors.affiliation: ADINACO Research Group, Department of Pharmaceutical Botany and Ecology, Faculty of Pharmacy, Charles University in Prague, Heyrovskeho 1203, Hradec Kralove 500 05, Czech Republic.
authors.forename: Lubomir
authors.initials: L
authors.lastname: Opletal
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, Pardubice 532 10, Czech Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Bilkova
mesh_terms: D047151:Amaryllidaceae Alkaloids / Q000494:pharmacology / Q000627:therapeutic use*; D000972:Antineoplastic Agents, Phytogenic / Q000032:analysis*; D017209:Apoptosis / Q000187:drug effects; D020169:Caspases / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D000071877:Checkpoint Kinase 1; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000378:metabolism; D004354:Drug Screening Assays, Antitumor; D016158:Genes, p53; D006801:Humans; D019169:Jurkat Cells; D007938:Leukemia / Q000188:drug therapy*; D019668:Liliaceae / Q000737:chemistry*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010617:Phenanthridines / Q000494:pharmacology / Q000627:therapeutic use*; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use; D011494:Protein Kinases / Q000378:metabolism
chemical_list: D047151:Amaryllidaceae Alkaloids; D000972:Antineoplastic Agents, Phytogenic; D019941:Cyclin-Dependent Kinase Inhibitor p16; D010617:Phenanthridines; D010936:Plant Extracts; C083906:hemanthidine; C462104:hemanthamine; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; D020169:Caspases
keywords: Apoptosis; Caspases; Cell cycle; Haemanthamine; Haemanthidine; Jurkat