Role for NF-kappa B in mediating the effects of hyperoxia on IGF-binding protein 2 promoter activity in lung alveolar epithelial cells.
The surface of the pulmonary alveolus is a major target for oxidant injury, and its proper repair following injury is dependent on the proliferative response of the stem cells of the alveolar epithelium, the type 2 cells. In previous studies on the mechanisms controlling this response, we have documented involvement of several components of the IGF system, and mainly of the IGF binding protein-2 (IGFBP-2). We have provided evidence that this binding protein was associated with inhibition of DNA synthesis of type 2 cells exposed to oxidants and that its expression was regulated mostly at the level of transcription. In the present study, we focused on the factors involved in this regulation. From examination of the IGFBP-2 gene promoter sequence which revealed the presence of four potential binding sites for transcription factors of the NF-kappa B/Rel family, we hypothesized that NF-kappa B might be involved in the transcriptional activation of IGFBP-2 in oxidant-exposed cells. Data reported herein demonstrated that NF-kappa B activated IGFBP-2 promoter in transient transfection assays, and that exposure of cells to hyperoxia was associated with accumulation of the active form of NF-kappa B. Using gel shift analysis, we documented in O2-treated cells an increased binding to the four NF-kappa B binding sites. We also showed that accumulation of NF-kappa B was associated with a decrease in the inhibitory molecule I kappa B-alpha. Based on the current knowledge on NF-kappa B regulation, it is likely that in a number of situations associated with injury of lung alveolar epithelial cells signaling events involving accumulation of NF-kappa B converge to activate IGFBP-2 and to block entry into S phase.
Cazals
V
V
Physiology Department-INSERM U142, Trousseau Hospital, St. Antoine Medical School, University of Paris, France.
Nabeyrat
E
E
Corroyer
S
S
de Keyzer
Y
Y
Clement
A
A
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002453:Cell Cycle; D002455:Cell Division; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D015342:DNA Probes / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism; D018496:Hyperoxia / Q000378:metabolism*; D018971:Insulin-Like Growth Factor Binding Protein 2 / Q000235:genetics*; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011650:Pulmonary Alveoli / Q000166:cytology / Q000293:injuries / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction; D015533:Transcriptional Activation; D014162:Transfection