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Effect of Amaryllidaceae alkaloids haemanthamine and haemanthidine on cell viability, apoptosis and cell cycle progression in human T-lymphoblast cell line
Authors: Havelek Radim | Seifrtová Martina | Královec Karel | Brůčková Lenka | Cahlíková Lucie | Řezáčová Martina | Bílková Zuzana
Year: 2013
Type of publication: ostatní - článek ve sborníku
Name of source: FEBS JOURNAL SUPPLEMENT
Publisher name: Wiley-Blackwell
Place: New York
Page from-to: 322-323
Titles:
Language Name Abstract Keywords
eng Effect of Amaryllidaceae alkaloids haemanthamine and haemanthidine on cell viability, apoptosis and cell cycle progression in human T-lymphoblast cell line Aim: To evaluate the effects of haemanthamine and haemanthidine, an isoquinoline alkaloids extracted from an Amaryllidaceae family plants, on human leukemic cells viability, cell cycle distributions and apoptosis. We also aim to clarify cellular and molecular mechanisms involved in the haemanthamine and haemanthidine-induced cell death in p53-null Jurkat cells. Methods: In this work, we measured the cytotoxic effects of micromolar concentrations of Amaryllidaceae alkaloids haemanthamine and haemanthidine on human leukemic Jurkat cells. Cell viability was determined using a dye-free system Casy Cell Counter. For apoptosis detection we used flow cytometry-based quantification of Annexin V-FITC and propidium iodide dual staining. Results were cross-verified using the fluorescence microscopy. Activity of caspases 3 was determined by Caspase-Glo3/7 luminescent assay. Dissipation of the mitochondrial membrane potential after treatments were monitored using JC-1. The cell cycle was analyzed by flow cytometry. Results: Our data showed a significant decrease in the percentage of viable cells after 24h treatment with both haemanthamine and haemanthidine. Furthermore, an increase in apoptosis was observed when cells were treated with both compounds. However, in haemanthidine-treated cells, there was a marked increase in Annexin V positive cells (cells that are in early apoptosis) contrary to haemanthamine-treated. Cells treated with both haemanthamine and haemanthidine for 24 hours had a reduction of the mitochondrial membrane potential. There was sharper decrease in the percentage of cells displaying an intact mitochondria after haemanthidine treatment compared to that after haemanthamine application. Haemanthamine and haemanthidine induced an increase in the percentages of cells in G1 and G2 cell cycle phase. Simultaneously these effects were accompanied by a decrease in S-phase cells.

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