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Suivi de la fonctionnalité des jonctions communicantes par la technique de gap-FRAP sur des modèles in vitro (2-D, 3-D) et ex vivo : intérêt pour le diagnostic du cancer

Abstract : Optical methods to characterize tissues for tumor detection are an area of diagnosis research. Gap-FRAP (Fluorescence recovery after photobleaching) technique is used to estimate gap junctions functionality. The majority of malignant cells are characterized by a modification of the number and functionality of gap junctions, compared to their healthy counterparts. Particularly used in vitro, we optimized the use of gap-FRAP technique for a potential transfer on tissues and organs ex vivo. We resolved using cell lines with a different connexin expression and distribution pattern, the choice of a fluorescent dye for further in vivo measurements: calcein-AM. The following stage consisted in developing a system engineering model for the comparative analysis of the fluorescence recovery data for bi-dimensional and three-dimensional (spheroid) models. The conditions previously defined have been transposed in entire organ, bladder rat. Multi-color staining was applied using a fluorescent dye to track to tumors cells in bladder ex vivo, a fluorescent marker to distinguish cellular details of the entire urothelium, and calcein-AM to assess GJIC. Gap-FRAP was used for the first time to differentiate intercellular communication degree between healthy and tumor tissue in entire organ ex vivo opening perspectives for the diagnosis of the bladder cancer correlated to feature modifications of GJIC.
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https://hal.univ-lorraine.fr/tel-01748453
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Muriel Abbaci. Suivi de la fonctionnalité des jonctions communicantes par la technique de gap-FRAP sur des modèles in vitro (2-D, 3-D) et ex vivo : intérêt pour le diagnostic du cancer. Autre. Université Henri Poincaré - Nancy 1, 2008. Français. ⟨NNT : 2008NAN10058⟩. ⟨tel-01748453⟩

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