title: CFTR impairment upregulates c-Src activity through IL-1β autocrine signaling.
abstract: Cystic Fibrosis (CF) is a disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Previously, we found several genes showing a differential expression in CFDE cells (epithelial cells derived from a CF patient). One corresponded to c-Src; its expression and activity was found increased in CFDE cells, acting as a signaling molecule between the CFTR activity and MUC1 overexpression. Here we report that bronchial IB3-1 cells (CF cells) also showed increased c-Src activity compared to 'CFTR-corrected' S9 cells. In addition, three different Caco-2 cell lines, each stably transfected with a different CFTR-specific shRNAs, displayed increased c-Src activity. The IL-1β receptor antagonist IL1RN reduced the c-Src activity of Caco-2/pRS26 cells (expressing a CFTR-specific shRNA). In addition, increased mitochondrial and cellular ROS levels were detected in Caco-2/pRS26 cells. ROS levels were partially reduced by incubation with PP2 (c-Src inhibitor) or IL1RN, and further reduced by using the NOX1/4 inhibitor GKT137831. Thus, IL-1β→c-Src and IL-1β→NOX signaling pathways appear to be responsible for the production of cellular and mitochondrial ROS in CFTR-KD cells. In conclusion, IL-1β constitutes a new step in the CFTR signaling pathway, located upstream of c-Src, which is stimulated in cells with impaired CFTR activity.
authors.affiliation: Laboratory of Cellular and Molecular Biology, Institute for Biomedical Research (BIOMED), School of Medical Sciences, Pontifical Catholic University of Argentina (UCA), National Scientific and Technical Research Council (CONICET), Buenos Aires, Argentina.
authors.forename: María Macarena
authors.initials: MM
authors.lastname: Massip-Copiz
authors.affiliation: Laboratory of Cellular and Molecular Biology, Institute for Biomedical Research (BIOMED), School of Medical Sciences, Pontifical Catholic University of Argentina (UCA), National Scientific and Technical Research Council (CONICET), Buenos Aires, Argentina.
authors.forename: Mariángeles
authors.initials: M
authors.lastname: Clauzure
authors.affiliation: Laboratory of Cellular and Molecular Biology, Institute for Biomedical Research (BIOMED), School of Medical Sciences, Pontifical Catholic University of Argentina (UCA), National Scientific and Technical Research Council (CONICET), Buenos Aires, Argentina.
authors.forename: Ángel Gabriel
authors.initials: ÁG
authors.lastname: Valdivieso
authors.affiliation: Laboratory of Cellular and Molecular Biology, Institute for Biomedical Research (BIOMED), School of Medical Sciences, Pontifical Catholic University of Argentina (UCA), National Scientific and Technical Research Council (CONICET), Buenos Aires, Argentina. Electronic address: tsantacoloma@gmail.com.
authors.forename: Tomás Antonio
authors.initials: TA
authors.lastname: Santa-Coloma
mesh_terms: D000818:Animals; D019898:Autocrine Communication; D000081247:CSK Tyrosine-Protein Kinase; D018938:Caco-2 Cells; D002460:Cell Line; D003550:Cystic Fibrosis / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000037:antagonists & inhibitors*; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism*; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism; D018396:Mucin-1 / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D061987:Sf9 Cells; D015398:Signal Transduction; D015854:Up-Regulation; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: C505032:CFTR protein, human; C508585:IL1B protein, human; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D053583:Interleukin-1beta; C060192:MUC1 protein, human; D018396:Mucin-1; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human
keywords: Autocrine loop; CFTR; Cystic fibrosis; DCFH-DA; IL-1β; IL1RN; MitoSOX; Mitochondria; Mitochondrial complex I; NOX; PP2; Reactive oxygen species (ROS); c-Src