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Apoptosis in tumour cells photosensitized with Rose Bengal acetate is induced by multiple organelle photodamage

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Abstract

Rose Bengal (RB) is a very efficient photosensitizer which undergoes inactivation of its photophysical and photochemical properties upon addition of a quencher group—i.e. acetate—to the xanthene rings. The resulting RB acetate (RB-Ac) derivative behaves as a fluorogenic substrate: it easily enters the cells where the native photoactive molecule is restored by esterase activities. It is known that the viability of RB-Ac-loaded cells is strongly reduced by light irradiation, attesting to the formation of intracellular RB. The aim of this study was to identify the organelles photodamaged by the intracellularly formed RB. RB-Ac preloaded rat C6 glioma cells and human HeLa cells were irradiated at 530 nm. Fluorescence confocal imaging and colocalization with specific dyes showed that the restored RB molecules redistribute dynamically through the cytoplasm, with the achievement of a dynamic equilibrium at 30 min after the administration, in the cell systems used; this accounted for a generalized damage to several organelles and cell structures (i.e. the endoplasmic reticulum, the Golgi apparatus, the mitochondria, and the cytoskeleton). The multiple organelle damage, furthermore, led preferentially to apoptosis as demonstrated by light and electron microscopy and by dual-fluorescence staining with FITC-labelled annexin V and propidium iodide.

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Acknowledgements

The authors wish to thank Ms Paola Veneroni for her technical assistance in cell culture. Dr Uliana De Simone is especially thanked for her assistance in enzyme histochemistry. Confocal fluorescence images were taken at the Centro Grandi Strumenti of the University of Pavia. This work was supported by the University of Pavia (FAR 2006) and MIUR (PRIN 2005058254).

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Authors and Affiliations

  1. Department of Animal Biology, University of Pavia, Piazza Botta 10, 27100, Pavia, Italy

    C. Soldani, M. G. Bottone, A. Fraschini, M. Biggiogera & C. Pellicciari

  2. Histochemistry and Cytometry Section, Institute of Molecular Genetics, National Research Council, Piazza Botta 10, 27100, Pavia, Italy

    A. C. Croce, M. G. Bottone, M. Biggiogera & G Bottiroli

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  1. C. Soldani
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  2. A. C. Croce
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  3. M. G. Bottone
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  4. A. Fraschini
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  5. M. Biggiogera
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  6. G Bottiroli
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  7. C. Pellicciari
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Correspondence to C. Pellicciari.

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Soldani, C., Croce, A.C., Bottone, M.G. et al. Apoptosis in tumour cells photosensitized with Rose Bengal acetate is induced by multiple organelle photodamage. Histochem Cell Biol 128, 485–495 (2007). https://doi.org/10.1007/s00418-007-0333-3

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  • DOI: https://doi.org/10.1007/s00418-007-0333-3

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