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

Maria Grazia Bottone, Cristiana Soldani, Annunzia Fraschini, Anna Cleta Croce, Giovanni Bottiroli, Tania Camboni, Anna Ivana Scovassi, Carlo Pellicciari

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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.

Original languageEnglish
Pages (from-to)391-399
Number of pages9
JournalHistochemistry and Cell Biology
Issue number3
Publication statusPublished - Mar 2009


  • Apoptosis
  • DNA electrophoresis
  • Fluorescence microscopy
  • Immunocytochemistry
  • Photosensitizers
  • Rose Bengal acetate

ASJC Scopus subject areas

  • Cell Biology
  • Histology
  • Medical Laboratory Technology
  • Molecular Biology


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