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Enzyme-assisted photosensitization activates different apoptotic pathways in Rose Bengal acetate treated HeLa cells

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Abstract

Photosensitization of tumor cells after incubation with Rose Bengal acetate (RB-Ac) induces multiple organelle photodamage followed by apoptotic cell death. We used immunocytochemical techniques in multicolor fluorescence microscopy to elucidate whether this occurs through the simultaneous activation of different apoptotic pathways, in HeLa cells. We detected in situ the activated forms of caspases 9 and 3, and the translocation from the mitochondria to the nucleus of the apoptosis inducing factor; DNA electrophoretic techniques were also used to assess the occurrence of nuclear DNA cleavage into either high- or low-molecular-weight fragments. Both the caspase-dependent and caspase-independent apoptotic pathways are activated. The genomic DNA is degraded into high molecular weight molecules only, without the formation of oligonucleosome-sized fragments. The ability of RB-Ac to induce the simultaneous release of apoptogenic signals from different photodamaged organelles makes it an especially powerful cytotoxic agent.

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Acknowledgments

We acknowledge the following Italian institutions for supporting our research: MIUR (PRIN project no. 2005058254), University of Pavia (FAR grant 2007) and Regione Lombardia (Project Metadistretti n. 4238). Confocal images were taken at the Centro Grandi Strumenti of the University of Pavia.

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Authors and Affiliations

  1. Dipartimento di Biologia Animale, Università degli Studi di Pavia, Piazza Botta, 10, 27100, Pavia, Italy

    Maria Grazia Bottone, Cristiana Soldani, Annunzia Fraschini & Carlo Pellicciari

  2. Istituto di Genetica Molecolare del CNR, Pavia, Italy

    Maria Grazia Bottone, Anna Cleta Croce, Giovanni Bottiroli, Tania Camboni & Anna Ivana Scovassi

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  1. Maria Grazia Bottone
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  2. Cristiana Soldani
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  3. Annunzia Fraschini
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  4. Anna Cleta Croce
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  5. Giovanni Bottiroli
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  6. Tania Camboni
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  7. Anna Ivana Scovassi
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  8. Carlo Pellicciari
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Correspondence to Maria Grazia Bottone.

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Bottone, M.G., Soldani, C., Fraschini, A. et al. Enzyme-assisted photosensitization activates different apoptotic pathways in Rose Bengal acetate treated HeLa cells. Histochem Cell Biol 131, 391–399 (2009). https://doi.org/10.1007/s00418-008-0538-0

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  • DOI: https://doi.org/10.1007/s00418-008-0538-0

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