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1 August 2000 Light-induced Apoptosis Involves a Defined Sequence of Cytoplasmic and Nuclear Calcium Release in AlPcS4-photosensitized Rat Bladder RR 1022 Epithelial Cells
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Abstract

Oxidative stress induced by light activation of photosensitizers is regarded to have a role in triggering cell death pathways during photodynamic therapy (PDT). Reactive oxygen species have been proposed to act as signal transduction molecules activating downstream reactions that lead to apoptosis. Mainly debated is the cooperating role of other signaling systems like calcium or pH. The present work contributes to this discussion by studying PDT effects in cell cultures of rat bladder epithelial cells for the hydrophilic tetrasulfonated aluminum phthalocyanine (AlPcS4). Cells were coincubated with the photosensitizer and the calcium-sensitive probe Fluo-3. The light-induced reactions were analyzed with a confocal laser scanning microscope. The dynamics of the process during light activation was observed with subcellular resolution. A transient calcium elevation during the irradiation process was detected, especially in the cell's nuclei, followed by a more sustained increase. The evaluation of the energy-dose–dependent phototoxicity after an incubation time with the photosensitizer of 1 and 24 h, showed enhanced phototoxicity when the drug was present for 24 h. Surprisingly, stimulation of cell proliferation was observed at very low light doses (at 0.2 J/cm2) when the drug was incubated for 24 h (cell viability 160%). Induction of apoptosis could be observed after irradiation with fluences between 1 and 3 J/cm2. Apoptotic cells were identified with fluorescein isothiocyanate–labeled Annexin V, which binds to phosphatidylserine after its translocation to the outer plasma membrane. In the presence of the antioxidant pyrrolidinedithiocarbamate the transient calcium elevation was totally inhibited, as was the subsequent translocation of PS. In contrast, N-acetyl-l-cysteine did not suppress the transient calcium increase. Our data might be consistent with calcium regulated processes during AlPcS4-PDT and the involvement of oxygen radicals.

Angelika Rück, Klaus Heckelsmiller, Roland Kaufmann, Nili Grossman, Elke Haseroth, and Nermin Akgün "Light-induced Apoptosis Involves a Defined Sequence of Cytoplasmic and Nuclear Calcium Release in AlPcS4-photosensitized Rat Bladder RR 1022 Epithelial Cells," Photochemistry and Photobiology 72(2), 210-216, (1 August 2000). https://doi.org/10.1562/0031-8655(2000)072<0210:LIAIAD>2.0.CO;2
Received: 26 November 1999; Accepted: 1 May 2000; Published: 1 August 2000
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