title: Retinoid signaling by all-trans retinoic acid and all-trans retinoyl-beta-D-glucuronide is attenuated by simultaneous exposure of human keratinocytes to retinol.
abstract: Retinol and retinyl esters are converted with time to slowly increasing amounts of all-trans retinoic acid (RA) in cultured human keratinocytes. Exogenous RA has been shown to limit retinol oxidation and to increase retinol esterification. Because significant amounts of retinol are present in biologic systems, we examined whether RA and all-trans-retinoyl-beta-D-glucuronide (RAG) interact with retinol in exhibiting their activities on HaCaT keratinocytes maintained in a retinoid-free culture system. RA was more potent than RAG and retinol in inducing ultrastructural changes attributed to retinoids, inhibiting cell proliferation as well as enhancing keratin 19 expression. In addition, retinoids were able to induce cellular retinoic acid-binding protein II mRNA levels in the cultures, whereas early RA and late RAG activity was detected. The described biologic effects of RA and RAG were diminished by simultaneous cell exposure to retinol. HaCaT cells quickly metabolized retinol to retinyl esters and consequently to low amounts of RA. RA treatment led to an early high peak of cellular RA followed by reduction to trace amounts. Treatment with RAG resulted in constantly high cellular RAG and low RA levels. Under the combined RA and retinol treatment retinyl esters were increased and RA was reduced in HaCaT cells, whereas extracellular RA levels were similar to those obtained by RA alone. On the other hand, the combination of RAG and retinol resulted in higher extracellular RAG, similar cellular RAG, and lower cellular RA levels than those obtained by RAG alone without any change in retinyl esters. This study demonstrates that retinoid signaling by RA and RAG is attenuated by simultaneous exposure of HaCaT keratinocytes in vitro to retinol. The presence of retinol in the medium alters the rate of RA or RAG metabolism and thus cellular RA concentrations. The intensity of retinoid signal is probably dependent on cellular RA levels. The resulting "antagonism" among retinoids is consistent with the presence of an auto-regulatory mechanism in human keratinocytes offering protection against excessive accumulation of cellular RA.
authors.affiliation: Department of Dermatology, University Medical Center Benjamin Franklin, The Free University of Berlin, Germany.
authors.forename: C C
authors.initials: CC
authors.lastname: Zouboulis
authors.forename: H
authors.initials: H
authors.lastname: Seltmann
authors.forename: J O
authors.initials: JO
authors.lastname: Sass
authors.forename: R
authors.initials: R
authors.lastname: Rühl
authors.forename: C
authors.initials: C
authors.lastname: Plum
authors.forename: U
authors.initials: U
authors.lastname: Hettmannsperger
authors.forename: U
authors.initials: U
authors.lastname: Blume-Peytavi
authors.forename: H
authors.initials: H
authors.lastname: Nau
authors.forename: C E
authors.initials: CE
authors.lastname: Orfanos
mesh_terms: D015152:Blotting, Northern; D002455:Cell Division; D002460:Cell Line / Q000166:cytology / Q000648:ultrastructure; D003470:Culture Media; D004347:Drug Interactions; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015603:Keratinocytes / Q000737:chemistry* / Q000187:drug effects / Q000502:physiology*; D007633:Keratins / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D018168:Receptors, Retinoic Acid / Q000235:genetics; D012176:Retinoids / Q000378:metabolism / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate; D014212:Tretinoin / Q000031:analogs & derivatives* / Q000502:physiology*; D014801:Vitamin A / Q000494:pharmacology*