Saltar al contenido
Merck

Glucocorticoids rapidly inhibit cell migration through a novel, non-transcriptional HDAC6 pathway.

Journal of cell science (2020-05-10)
Stephen Kershaw, David J Morgan, James Boyd, David G Spiller, Gareth Kitchen, Egor Zindy, Mudassar Iqbal, Magnus Rattray, Christopher M Sanderson, Andrew Brass, Claus Jorgensen, Tracy Hussell, Laura C Matthews, David W Ray
RESUMEN

Glucocorticoids (GCs) act through the glucocorticoid receptor (GR, also known as NR3C1) to regulate immunity, energy metabolism and tissue repair. Upon ligand binding, activated GR mediates cellular effects by regulating gene expression, but some GR effects can occur rapidly without new transcription. Here, we show that GCs rapidly inhibit cell migration, in response to both GR agonist and antagonist ligand binding. The inhibitory effect on migration is prevented by GR knockdown with siRNA, confirming GR specificity, but not by actinomycin D treatment, suggesting a non-transcriptional mechanism. We identified a rapid onset increase in microtubule polymerisation following GC treatment, identifying cytoskeletal stabilisation as the likely mechanism of action. HDAC6 overexpression, but not knockdown of αTAT1, rescued the GC effect, implicating HDAC6 as the GR effector. Consistent with this hypothesis, ligand-dependent cytoplasmic interaction between GR and HDAC6 was demonstrated by quantitative imaging. Taken together, we propose that activated GR inhibits HDAC6 function, and thereby increases the stability of the microtubule network to reduce cell motility. We therefore report a novel, non-transcriptional mechanism whereby GCs impair cell motility through inhibition of HDAC6 and rapid reorganization of the cell architecture.This article has an associated First Person interview with the first author of the paper.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Dimetilsulfóxido, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
Sigma-Aldrich
Dexametasona, powder, BioReagent, suitable for cell culture, ≥97%
Sigma-Aldrich
Phosphatase Inhibitor Cocktail 2, aqueous solution (dark coloration may develop upon storage, which does not affect the activity)
Sigma-Aldrich
Cóctel de inhibidores de fosfatasa 3, DMSO solution
Sigma-Aldrich
Anti-α-tubulina monoclonal antibody produced in mouse, ascites fluid, clone B-5-1-2
Sigma-Aldrich
Mifepristone, ≥98%
Sigma-Aldrich
Trichostatin A, ≥98% (HPLC), from Streptomyces sp.
Sigma-Aldrich
Nicotinamide, ≥98% (HPLC), powder
Sigma-Aldrich
bisBenzimide H 33342 trihydrochloride, for fluorescence, ≥97.0% (HPLC)
Sigma-Aldrich
Fluticasone propionate, ≥98% (HPLC), powder
Sigma-Aldrich
Tubacin, ≥98% (HPLC)
Sigma-Aldrich
Anti-ATAT1 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution