Développement d'outils « biocapteurs/modèle cellulaire » pour l'identification de ligands et l'étude des interactions ADN/protéine et protéines/protéines

摘要

Estrogens could modulate gene expression by interacting with Estrogens Receptors (ER). These receptors were described as transcription factor and are able to interact with a specific DNA sequence called Estrogens Response Element (ERE). The screening of molecules able to link to ER and to modulate gene expression could be applied to new therapeutic developments (menopauses, hormonodependant cancers) or could have an impact on healthcare and environment protection (endocrine disturbers). We have designed two Surface Plasmons Resonance (SPR) biosensors to develop a fast and low cost molecular screening strategy. In order to validate the routine application of these tools, SPR results must be correlated with more classical data obtained with a cellular model. That was why we have transfected a human breast cancer cell line (MCF-7) with a reporter vector. We have identified a new ERa agonist phytoestrogene (7-O-ß-D-glucopyranolychrysin) using this double approach. Furthermore, one of ours biosensors was also applicable to the proteins/proteins interactions detection. We have decided to use this sensor to identify new partners of GEC1 protein. In conclusion, we have designed in parallel DNA/protein biosensors and cellular model in order to identify xenoestrogen (7-O-ß-D-glucopyranolychrysin). We have also developed a proteins/proteins biosensor which was consistent with protein partner identification.