Genetically encoded photosensitizers are a unique instrument for investigation of cellular mechanisms of photodynamic therapy (PDT). Fluorescent flavoprotein miniSOG (mini Singlet Oxygen Generator) generates singlet oxygen with a high yield and demonstrates strong phototoxic properties in vitro in cancer cells. However, the effective approaches for PDT with miniSOG have not been developed so far. The purpose of the study is to investigate phototoxic effects of miniSOG induced by continuous wave (CW) or pulsed laser irradiation in tumor spheroids. We found that maximum photobleaching of miniSOG without temperature effects was achieved at 120 mW/cm2 in CW mode or pulse periodic mode. PDT in pulse periodic mode provided more pronounced increase in the number of dead cells in comparison with CW mode and, moreover, induced apoptosis more efficiently. Therefore, we report for the first time on an effective regimen for PDT with miniSOG in a tumor spheroid model using pulsed periodic laser irradiation.
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