Characterization of the anti-Leishmania effect induced by cisplatin, an anticancer drug.
The cis-diamminedichloroplatinum(II), known as cis-DDP or cisplatin is a widely used drug in cancer chemotherapy. Although a recent study has shown the anti-Leishmania activity of some cis-DDP derivatives, the cytotoxic properties were measured only on promastigotes, the insect vector form of the parasite. In this study the effect of cis-DDP on promastigotes and amastigotes, the vertebrate stage of the parasite is reported. The IC50, determined by flow cytometry, after 72 h of drug incubation was four times higher, 7.73+/-1.03 microM in the case of promastigotes compared to axenic amastigotes, 1.88+/-0.10 microM. In intracellular amastigotes the IC50, determined by counting the parasite index was 1.85+/-0.22 microM. By using flow cytometry, two patterns of cell cycle changes was observed: cis-DDP treated promastigotes and amastigotes accumulated in S phase and G2 phase, respectively. The cis-DDP response was also found to involve an "apoptosis-like" death of both promastigotes and amastigotes. However, DNA fragmentation was only detected in promastigote forms. In contrast mitochondrial transmembrane potential loss was observed for both stages of the parasite. Upon incubation of parasites with the drug an increase on GSH and GSSG levels and reactive oxygen species could be detected in the case of promastigote. Moreover, a slight increase of GSH level was detected on amastigote form. Taken together, these observations indicate that amastigotes are more sensitive to cis-DDP when compared to promastigotes. However, the signaling pathways leading to cell death could be different.
Laboratório de Bioquímica, Faculdade de Farmácia da Universidade do Porto, Porto, Portugal.
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Tavares
M
M
Ouaissi
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Ouaissi
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A
Cordeiro-da-Silva
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000502:physiology; D002945:Cisplatin / Q000494:pharmacology*; D016054:DNA, Protozoan / Q000187:drug effects / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D019803:Glutathione Disulfide / Q000378:metabolism; D020287:In Situ Nick-End Labeling / Q000379:methods; D018314:Leishmania infantum / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D007898:Leishmaniasis, Visceral / Q000469:parasitology*; D008018:Life Cycle Stages; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010718:Phosphatidylserines / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
D000970:Antineoplastic Agents; D016054:DNA, Protozoan; D010718:Phosphatidylserines; D017382:Reactive Oxygen Species; D005978:Glutathione; D002945:Cisplatin; D019803:Glutathione Disulfide