Photolon™ - photosensitization induces apoptosis via ros-mediated cross-talk between mitochondria and lysosomes

Abstract

The localization of photosensitizers in the subcellular compartments during photodynamic therapy (PDT) plays a major role in the cell destruction; therefore, the aim of this study was to investigate the intracellular localization of Chlorin e6-PVP (Photolon (TM)) in malignant and normal cells. Our study involves the characterization of the structural determinants of subcellular localization of Photo Ion, and how subcellular localization affects the selective toxicity of Photo Ion towards tumor cells. Using confocal laser scanning microscopy (CLSM) and fluorescent organelle probes; we examined the subcellular localization of Photolon (TM) in the murine colon carcinoma CT-26 and normal fibroblast (NHLC) cells. Our results demonstrated that after 30 min of incubation, the distribution of Photo Ion was localized mainly in the cytoplasmic organelles including the mitochondria, lysosomes, Golgi apparatus, around the nuclear envelope and also in the nucleus but not in the endoplasmic reticulum whereas in NHLC cells, Photo Ion was found to be localized minimally only in the nucleus not in other organelles studied. The relationship between subcellular localization of Photo Ion and PDT-induced apoptosis was investigated. Apoptotic cell death was judged by the formation of known apoptotic hallmarks including, the phosphatidylserine externalization (PS), PARP cleavage, a substrate for caspase-3 and the formation of apoptotic nuclei. At the irradiation dose of 1 J/cm(2), the percentage of apoptotic cells was 80%, respectively. This study provided substantial evidence that Photo Ion preferentially localized in the subcellular organelles in the following order: nucleus, mitochondria, lysosomes and the Golgi apparatus and subsequent photodamage of the mitochondria and lysosomes played an important role in PDT-mediated apoptosis CT-26 cells. Our results based on the cytoplasmic organelles and the intranuclear localization extensively enhance the efficacy of PDT with appropriate photosensitizer and light dose and support the idea that PDT can contribute to elimination of malignant cells by inducing apoptosis, which is of physiological significance.