Steroid responsiveness of renal epithelial cells. Dissociation of transrepression and transactivation.
Glucocorticoids modulate cellular and inflammatory responses via stimulation or inhibition of gene transcription. Inhibition of cytokine gene expression is mediated via repression of transcription factors, including NF-kappaB. Previously we have shown that cytokine production by renal epithelial cells is insensitive to the inhibitory action of dexamethasone. In this study we demonstrate that dexamethasone is unable to inhibit NF-kappaB activation in the renal epithelial cell line HK-2, as measured by IkappaB-alpha degradation and DNA binding activity. Transfection of an NF-kappaB-inducible reporter gene demonstrated that non-stimulated HK-2 cells contain a high level of constitutively active NF-kappaB compared with the steroid-sensitive airway epithelial cell line A549, which was not blocked by dexamethasone. Expression and nuclear translocation of the glucocorticoid receptor (GR) was comparable in both cell types. In HK-2 cells, dexamethasone stimulated expression of two glucocorticoid-responsive genes, beta(2)-adrenoreceptors and angiotensinogen. The capacity of GR to transactivate the native angiotensinogen glucocorticoid-responsive element (GRE) using chromatin-IP was not impaired. Moreover, dexamethasone activation of a GRE-driven reporter construct appeared to be equally effective, although less sensitive compared with A549 cells. In conclusion, we provide evidence that glucocorticoids are unable to repress the activity of NF-kappaB in renal epithelial cells in the presence of an intact stimulatory pathway.
de Haij
Simone
S
Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.
Adcock
Ian M
IM
Bakker
Astrid C
AC
Gobin
Sam J P
SJ
Daha
Mohamed R
MR
van Kooten
Cees
C
D002460:Cell Line; D003907:Dexamethasone / Q000378:metabolism* / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism*; D005938:Glucocorticoids / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects
D005938:Glucocorticoids; D020672:I-kappa B Proteins; D016328:NF-kappa B; D003907:Dexamethasone