Analyse par spectrométrie de masse de l'oxygène moléculaire singulet et de protéines potentiellement ciblées au sein de cellules tumorales lors de la thérapie photodynamique

Abstract : Photodynamic therapy (PDT), uses a photosensitizing molecule such as 5,10,15,20-tetrakis(m-hydroxyphenyl) chlorin (m-THPC, Foscan®), a second generation drug which is specially targeted tumoural tissue with a good selectivity, light and oxygen, inducing cell death by necrosis or apoptosis. Firstly, our studies consist to detect by MALDI-TOF/MS, in intact HT29 cells (adenocarcinoma human colon), singlet oxygen generated by Foscan® (Biolitec Pharma Ldt, Dublin, Irlande) and the protein cells distribution after PDT treatment. A MALDI-TOF mass spectrometer was used to highlight ortho-benzoïbenzene (o-BB) resulting from the reaction between singlet oxygen generated by Foscan® during PDT treatment and 1,3-diphenylisobenzofurane (1,3-DPBF, a specific singlet oxygen quencher). This technique allows the following of the in situ behaviour of the photosensitizer and to detect the presence of singlet oxygen directly in intact HT29 cells. We have also studied the oxidative stress induced by PDT treatment on HT29 cells. After 2D gel SDS-PAGE step in order to observe the protein distribution, proteomic approach is carried out by MALDI-FTICR mass spectrometry (9,4 T, Ion Spec Varian, California). Thnaks to ImageMaster 2D platinium software, we are able to visualize under expression of some proteins. The proteinic finger printings is then characterized by MALDI-FT-ICR/MS and the first results indicate that proteins of dislfide Isomerase family should be implied in PDT processes. MALDI-TOF/MS and MALDI-FTICRMS (9,4 T) appear to be a sensitive and reliable analytical tool (add to UV/Visible anf fluorescence spectroscopy) for the mechanism of PDT understanding
Mots-clés : Photochimiothérapie
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Marc Dodeller. Analyse par spectrométrie de masse de l'oxygène moléculaire singulet et de protéines potentiellement ciblées au sein de cellules tumorales lors de la thérapie photodynamique. Chimie thérapeutique. Université Paul Verlaine - Metz, 2007. Français. ⟨NNT : 2007METZ053S⟩. ⟨tel-01775420⟩

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