Involvement of mitochondria and ROS generation in edelfosine-induced cell death in <i>Leishmania</i> parasites and cancer cells.
Janny A. Villa-Pulgarín
Consuelo Gajate
Javier Botet
Alberto Jimenez
Nicole Justies
Rubén E. Varela-M
Álvaro Cuesta-Marbán
Ingrid Müller
Manuel Modolell
José L. Revuelta
Faustino Mollinedo
10.1371/journal.pntd.0005805.g005
https://plos.figshare.com/articles/figure/Involvement_of_mitochondria_and_ROS_generation_in_edelfosine-induced_cell_death_in_i_Leishmania_i_parasites_and_cancer_cells_/5332339
<p>(<b>A</b>) <i>L</i>. <i>panamensis</i> promastigotes were untreated (Control) or treated with 10 μM edelfosine at the indicated times, and cells with disrupted ΔΨ<sub>m</sub> (DiOC<sub>6</sub>(3)<sup>low</sup>) and ROS production (HE→Eth) were measured by flow cytometry. The numbers in each quadrant refer to the percentages of cells in each population. (<b>B</b>) <i>L</i>. <i>panamensis</i> promastigotes untreated (Control) and treated with edelfosine (EDLF) for 3 h were incubated with 2 μM HE and 10 μg/ml Hoechst 33342, and then analyzed by fluorescence microscopy. (<b>C</b>) <i>L</i>. <i>panamensis</i> promastigotes and (<b>D</b>) Jurkat cells were preincubated with 10 μg/ml CsA for 1 h, or with 10 mM NAC or 10 mM GSH for 2 h, and then incubated in the absence or presence of 10 μM edelfosine for 9 h. After treatment, the percentage of hypodiploid cells was analyzed by flow cytometry. Untreated control cells were run in parallel. (<b>E</b>) <i>L</i>. <i>panamensis</i> promastigotes and (<b>F</b>) Jurkat cells were preincubated with 10 μM rotenone, 5 mM malonate, 10 μM antimycin A, 1.5 mM azide, 50 μM CCCP or with 1 and 10 μM oligomycin (<i>L</i>. <i>panamensis</i> and Jurkat cells, respectively) for 1 h, and then incubated in the absence or presence of 10 μM edelfosine for 9 h. After treatment, cells producing ROS were quantified by flow cytometry. Untreated control cells were run in parallel. Data shown are means ± SD or representative of three independent experiments performed. Asterisks denote that the differences between the indicated groups (C and D) and with control cells (E and F) are statistically significant. (*) <i>P</i><0.05. (**) <i>P</i><0.01.</p>
2017-08-22 17:26:14
ATP synthase
lipid raft-located F O F 1
F O F 1
ATP synthase inhibition
Saccharomyces cerevisiae yeast
APL
tumor cells
tumor cell apoptosis-inducer edelfosine
edelfosine drug resistance
Leishmania axenic amastigotes
ether lipid edelfosine Background Leishmaniasis
DNA
novel druggable targets
Leishmania parasites
raft-located F O F 1
vivo antileishmanial activity
tumor cell mitochondria
edelfosine-induced cell death
intracellular Leishmania amastigotes
show antileishmanial activity
macrophage pro-inflammatory responses