Gas chromatography/mass spectrometry characterization of corticosteroid metabolism in human immortalized keratinocytes
Open accessAbstract
To continue our studies on the cutaneous expression of a proopiomelanocortin/corticotropin-releasing hormone system, we investigated whether this is accompanied by adrenal-type enzymatic activity. Immortalized cultured human keratinocytes were incubated with radiolabeled corticosteroids. Analysis by thin-layer chromatography showed rapid transformation of both progesterone and deoxycorticosterone; one of the progesterone metabolites migrated at the same rate as deoxycorticosterone. Gas chromatography/mass spectrometry further identified as major species of deoxycorticosterone metabolites 3beta,6alpha,21-trihydroxy-5alpha-pregnan-20-one, 3alpha,6alpha,21-trihydroxy-5alpha-pregnan-20-one, and 3alpha5alpha- and 3beta5alpha-tetrahydrodeoxycorticosterone. Minor metabolites were 3alpha,21-dihydroxy-5-pregnen-20- one (3alphaDelta5-21-OHpregnenolone), 3beta,21-dihydroxy-5-pregnen-20-one (3betaDelta5-21-OHpregnenolone), 3alpha,21-dihydroxy-4-pregnen-20-one (3alphaDelta4-21-OHpregnenolone), 6-hydroxy-dihydrodeoxycorticosterone, and two 5-dihydrodeoxycorticosterone species. Thus, in addition to sex steroids keratinocytes also actively metabolize corticosteroids along similar enzymatic pathways. The surprising detection of 3alphaDelta5-21-OHpregnenolone and 3 betaDelta5-21-OHpregnenolone, indicating Delta4-ketosteroids to Delta5-hydroxysteroids conversion, provides strong evidence for the occurrence, at least in human keratinocytes, of isomerase activity that allows the reaction to proceed in reverse of its usual direction. As skin expresses 3alpha/beta-hydroxysteroid dehydrogenase/Delta5-Delta4 isomerases, cutaneous reactions catalyzed by these enzymes must be reversible. In conclusion, besides elements of the corticotropin-releasing hormone/proopiomelanocortin system human keratinocytes show high levels of corticosteroid metabolizing activity. Moreover, the wide array of steroid products generated from a single substrate indicates serial progressive conversion involving 5alpha-reductase, 6alpha-hydroxylase, 3alpha/beta-hydroxysteroid dehydrogenase, and reverse Delta4minus signDelta5 isomerase enzymes. As distinct from the adrenal cortex, production of A, B, Aldo, 18OHdeoxycorticosterone, or F in keratinocytes was absent or below limits of detectability.
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In HaCaT keratinocytes, progesterone is rapidly metabolized to DOC and then to several products other than corticosterone, aldosterone, and cortisol, including 3β,6α,21-trihydroxy-5α-pregnan-20-one, 3α,6α,21-trihydroxy-α-pregnan-20-one, and 3α,5α- and 3β,5α-tetrahydrodeoxycorticosterone.
“In our studies on HaCaT keratinocytes, we have detected rapid metabolism of progesterone to DOC with further transformation to several products different from corticosterone, aldosterone and cortisol, some of which were identified by GC/MS as 3β,6α,21-trihydroxy-5α-pregnan-20-one, 3α,6α,21-trihydroxy-α-pregnan-20-one, and 3α,5α-and 3β,5α-tetrahydrodeoxycorticosterone [80].”
From Steroidogenesis in the skin: Implications for local immune functions (2013) · cites this paper as [80]
The low rate of cutaneous steroid production can explain why earlier studies did not detect cortisol formation by keratinocytes.
“This low rate of production could explain the lack of detection of cortisol formation by keratinocytes in earlier studies by Milevich et al. [78] and later by us [79].”
From Steroidogenesis in the skin: Implications for local immune functions (2013) · cites this paper as [79]
In Smith-Lemli-Opitz syndrome, deficiency of 7-dehydrocholesterol reductase leads to UVA photosensitivity of the skin and to formation of atypical steroids.
“Firstly, the 7-Δ reductase deficiency observed in Smith–Lemli–Opitz syndrome (SLOS), in an addition to severe malformations, results in development of skin photosensitivity to UVA [158, 159] and production a series of unusual steroids [79, 160, 161].”
From Steroidogenesis in the skin: Implications for local immune functions (2013) · cites this paper as [79]
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References 33 · Cited by 41
- Authors
- Slominski, A., Wortsman, J., Foecking, M. F., Shackleton, C., Gomez-Sanchez, C., & Szczesniewski, A.
- Affiliations
- University of Tennessee Health Science Center (US)
- SIU Medicine (US)
- Harry S. Truman Memorial Veterans' Hospital (US)
- University of Mississippi Medical Center (US)
- Corresponding author
- Slominski, Andrzej
- ORCID
- Slominski, Andrzej
- Venue
- Journal of Investigative Dermatology
- Publisher
- Elsevier BV
- Published
- 2002-02
- Volume
- 118
- Issue
- 2
- Pages
- 310-315
- ISSN
- 0022-202X (ISSN-L)
- OpenAlex
- W1979039462
- Cited by
- 41
- References
- 33
- Citations by year
- 2025: 1 · 2023: 1 · 2022: 1 · 2021: 1 · 2019: 1 · 2017: 1 · 2016: 3 · 2014: 3 · 2013: 2 · 2012: 4
- FWCI
- 1.71
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Journal of Investigative Dermatology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (HL-27737, R01HL027255, HL27255, R01HL027737)
- Topics
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Steroid Chemistry and Biochemistry
- Keywords
- corticosteroid metabolism, human keratinocytes, deoxycorticosterone, progesterone
- MeSH terms
- Cell Line, Transformed, Chromatography, High Pressure Liquid, Chromatography, Thin Layer, Desoxycorticosterone, Humans, Gas Chromatography-Mass Spectrometry, Progesterone, Keratinocytes
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