NF-kappaB activation is involved in regulation of cystic fibrosis transmembrane conductance regulator (CFTR) by interleukin-1beta.
Interleukin-1 beta (IL-1beta) regulates the levels of cystic fibrosis transmembrane conductance regulator (CFTR) mRNA and protein in the T84 human carcinoma cell line. Here, we studied the role of the transcription factor NF-kappaB in this regulation. Initially, T84 cells were pretreated with the NF-kappaB inhibitor pyrrolidine dithiocarbamate. Cells were then stimulated with IL-1beta, and CFTR mRNA levels were determined after 4 h by Northern blot analysis. As a result of PDTC treatment, IL-1beta stimulation of CFTR mRNA was blocked. On the other hand, daunorubicin, an NF-kappaB activator, increased the steady-state levels of CFTR mRNA. Furthermore, after treatment with IL-1beta for 1 h, cytoplasmic IkappaBalpha degradation occurred simultaneously with translocation of p65 into the nucleus. The T84 cells were also transduced with an adenoviral vector expressing a dominant negative form of IkappaBalpha, which prevents IkappaBalpha phosphorylation and the subsequent nuclear translocation of NF-kappaB. After viral transduction, the cells were stimulated with IL-1beta for 4 h, and CFTR mRNA levels were measured by Northern blot analysis. The stimulation of CFTR, induced by IL-1beta, was also blocked in the presence of the dominant negative mutant. These results indicate that NF-kappaB is involved in the pathway by which IL-1beta regulates CFTR.
Instituto de Investigaciones Bioquímicas-Fundación Campomar (IIB, UBA; IIBBA, CONICET), 1405 Buenos Aires, Argentina.
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D001483:Base Sequence; D015153:Blotting, Western; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D016328:NF-kappa B / Q000378:metabolism*; D015345:Oligonucleotide Probes; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000187:drug effects
C505032:CFTR protein, human; D007375:Interleukin-1; D016328:NF-kappa B; D015345:Oligonucleotide Probes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator