Restoration of ENaC expression by glucocorticoid receptor transfection in human HT-29/B6 colon cells

Biochemical and Biophysical Research Communications
Sebastian ZeissigJörg D Schulzke

Abstract

The epithelial sodium channel (ENaC) controls colonic sodium absorption. So far, investigation of ENaC was limited by an unexplained lack of steroid-dependent ENaC expression in cultured intestinal cells, which we aimed to resolve. HT-29/B6 cells constitutively expressed the alpha-ENaC subunit, while beta- and gamma-ENaC subunits could not be detected due to deficient basal as well as corticosteroid-induced transcription. This was due to a lack of expression of both activating and inhibiting isoforms of glucocorticoid receptor (GR-alpha, -beta) and mineralocorticoid receptor. Stable transfection of GR-alpha restored intestine-specific glucocorticoid upregulation of beta- and gamma-ENaC in HT-29/B6 cells, which was followed by intact targeting of ENaC channels to the apical cell membrane and dose-dependent induction of electrogenic sodium absorption. In conclusion, ENaC deficiency is due to a lack of steroid receptors and not the consequence of a crypt-like phenotype of cultured intestinal cells. By stable GR transfection we obtained a model, in which ENaC regulation can be studied.

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Citations

Nov 13, 2008·Biochemical and Biophysical Research Communications·Salah AmashehMichael Fromm
Mar 12, 2009·Biochemical and Biophysical Research Communications·Theresa BergannJörg D Schulzke
Oct 17, 2015·American Journal of Respiratory Cell and Molecular Biology·Felix KielgastOliver H Wittekindt

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