rnai-based screening identifies kinases interfering with dioxin-mediated up-regulation of cyp1a1 activityrnai-based筛选识别激酶干扰dioxin-mediated老年病的cyp1a1活动.pdfVIP

RNAi-Based Screening Identifies Kinases Interfering with Dioxin-Mediated Up-Regulation of CYP1A1 Activity 1 ¤ 1 1 ´ 1 1 David Gilot * , Nolwenn Le Meur , Fanny Giudicelli , Marc Le Vee , Dominique Lagadic-Gossmann , ´ 1, Olivier Fardel1,2 Nathalie Theret ´ ´ ´ ´ ´ 1 EA 4427 Signalisation et Reponse aux Agents Infectieux et Chimiques, Universite de Rennes 1, Institut de Recherche Sante, Environnement et Travail; Institut Federatif de ´ ´ ´ Recherche 140, Rennes, France, 2 Departement Hematologie, Immunologie et Therapie Cellulaire, Hopital Pontchaillou, CHU Rennes, Rennes, France Abstract Background: The aryl hydrocarbon receptor (AhR) is a transcription factor activated by several environmental pollutants, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and involved in carcinogenesis and various physiological processes, including immune response and endocrine functions. Characterization of kinases-related AhR transduction pathway remains an important purpose. Results: We performed a kinome-wide siRNA screen in human mammary MCF-7 cells to identify non redundant protein kinases implicated in the up-regulation of cytochrome P-450 (CYP) 1A1 activity, an AhR referent target, in response to TCDD exposure. To this aim, we monitored CYP1A1-related ethoxyresorufin-O-deethylase (EROD) activity and quantified cell density. This normalization was crucial since it allowed us to focus only on siRNA affecting EROD activity and discard siRNA affecting