Download PDF
Horm Metab Res 2007; 39(2): 141-148
DOI: 10.1055/s-2007-961814
Original

© Georg Thieme Verlag KG Stuttgart · New York

Inhibition of 5α-Reductase Activity in SZ95 Sebocytes and HaCaT Keratinocytes In Vitro

K. Seiffert 1 , H. Seltmann 2 , M. Fritsch 3 , C. C. Zouboulis 2 , 4
  • 1Division of Dermatology and Cutaneous Sciences, Michigan State University, East Lansing MI, USA
  • 2Laboratory of Biogerontology, Dermato-Pharmacology, and Dermato-Endocrinology, Institute of Clinical Pharmacology and Toxicology, Charité Universitaetsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany
  • 3Schering Research Laboratories, Schering AG, Berlin, Germany
  • 4Departments of Dermatology and Immunology, Dessau Medical Center, Dessau, Germany
Further Information

Publication History

received 5. 9. 2006

accepted 8. 11. 2006

Publication Date:
27 February 2007 (online)

Also available at
    Permissions and Reprints

Abstract

Inhibition of 5α-reductase type 1 has been considered to be a promising target for treatment of androgen-dependent skin disorders, however, currently published clinical results on acne treatment are rather disappointing. In this study, the influence of selective inhibitors of 5α-reductase on testosterone metabolism within SZ95 sebocytes and HaCaT keratinocytes in vitro was investigated. In both cell types, the isotype 1 inhibitor MK386 completely inhibited the conversion of testosterone to 5α-dihydrotestosterone in concentrations higher than 10-9 M. Inhibitors of the isotype 2 such as finasteride, dihydrofinasteride, and turosteride, were >100-fold less active, while, as expected, androgen receptor inhibitors did not affect the 5α-reductase activity. MK386, but not finasteride, reduced testosterone-stimulated proliferation and slightly reduced the testosterone-induced increase in the amount of SZ95 sebocyte proteins. The androgen receptor inhibitor cyproterone acetate exhibited no effect on testosterone-induced proliferation, but inhibited the 5α-dihydrotestosterone-induced sebocyte proliferation. Our experimental findings and the existing clinical results indicate that the inhibition of 5α-reductase activity alone may be insufficient to reduce overall sebocyte activity and improve acne lesions.

Keywords

androgens - anti-androgens - 5α-reductase inhibitors - sebaceous gland - keratinocytes - cell proliferation - acne

References

  • 1 Chen W, Thiboutot D, Zouboulis CC. Cutaneous androgen metabolism - Basic research and clinical perspectives.  J Invest Dermatol. 2002;  119 992-1007
    CrossrefPubMedSearch in Google Scholar
  • 2 Zouboulis CC, Degitz K. Androgen action on human skin - From basic research to clinical significance.  Exp Dermatol. 2004;  13 ((Suppl 4)) 5-10
    PubMedSearch in Google Scholar
  • 3 Sansone G, Reisner RM. Differential rates of conversion of testosterone to dihydrotestosterone in acne and in normal human skin - a possible pathogenic factor in acne.  J Invest Dermatol. 1971;  56 36-72
    PubMedSearch in Google Scholar
  • 4 Chen W, Yang C-C, Sheu H-M, Seltmann H, Zouboulis CC. Expression of sex-determining genes in human sebaceous glands and their possible roles in pathogenesis of acne.  J Eur Acad Dermatol Venereol. , in press
    PubMed
  • 5 Chen W, Tsai S-J, Liao C-Y, Tsai R-Y, Chen Y-R, Pan B-J, Hung C-L, Zouboulis CC. Higher levels of steroidogenic acute regulatory protein and type I 3β-hydroxysteroid dehydrogenase in the scalp of men with androgenetic alopecia.  J Invest Dermatol. 2006;  126 2332-2335
    CrossrefPubMedSearch in Google Scholar
  • 6 Thiboutot D, Harris G, Iles V, Cimis G, Guilliland H, Hagari S. Activity of the type 1 5alpha-reductase exhibits regional differences in isolated sebaceous glands and whole skin.  J Invest Dermatol. 1995;  105 209-214
    CrossrefPubMedSearch in Google Scholar
  • 7 Chen W, Zouboulis CC, Fritsch M, Blume-Peytavi U, Kodelja V, Goerdt S, Luu-The V, Orfanos CE. Evidence of heterogeneity and quantitative differences of the type 1 5α-reductase expression in cultured human skin cells. Evidence of its presence in melanocytes.  J Invest Dermatol. 1998;  110 84-89
    CrossrefPubMedSearch in Google Scholar
  • 8 Luu-The V, Sugimoto Y, Puy L, Labrie Y, Lopez Solache I, Singh M, Labrie F. Characterization, expression and immunohistochemical localization of 5 alpha-reductase in human skin.  J Invest Dermatol. 1994;  102 221-226
    CrossrefPubMedSearch in Google Scholar
  • 9 Eicheler W, Dreher M, Hoffmann R, Happle R, Aumüller G. Immunohistochemical evidence for differential distribution of 5 alpha-reductase isoenzymes in human skin.  Br J Dermatol. 1995;  133 371-376
    CrossrefPubMedSearch in Google Scholar
  • 10 Bayne EK, Flanagan J, Einstein M, Ayala J, Chang B, Azzolina B, Whiting DA, Mumford RA, Thiboutot D, Singer II, Harris G. Immunohistochemical localization of types 1 and 2 5α-reductase in human scalp.  Br J Dermatol. 1999;  141 481-491
    CrossrefPubMedSearch in Google Scholar
  • 11 Thiboutot D, Bayne E, Thorne J, Gilliland K, Flanagan J, Shao Q, Light J, Helm K. Immunolocalization of 5alpha-reductase isozymes in acne lesions and normal skin.  Arch Dermatol. 2000;  136 1125-1129
    PubMedSearch in Google Scholar
  • 12 Fritsch M, Orfanos CE, Zouboulis CC. Sebocytes are the key regulators of androgen homeostasis in human skin.  J Invest Dermatol. 2001;  116 793-800
    CrossrefPubMedSearch in Google Scholar
  • 13 Leyden J, Bergfeld W, Drake L, Dunlap F, Goldman MP, Gottlieb AB, Heffernan MP, Hickman JG, Hordinsky M, Jarrett M, Kang S, Lucky A, Peck G, Phillips T, Rapaport M, Roberts J, Savin R, Sawaya ME, Shalita A, Shavin J, Shaw JC, Stein L, Stewart D, Strauss J, Swinehart J, Swinyer L, Thiboutot D, Washenik K, Weinstein G, Whiting D, Pappas F, Sanchez M, Terranella L, Waldstreicher J. A systemic type I 5 alpha-reductase inhibitor is ineffective in the treatment of acne vulgaris.  J Am Acad Dermatol. 2004;  50 443-447
    CrossrefPubMedSearch in Google Scholar
  • 14 Bowring N. The development of BRL 7660 as an anti-acne agent.  Skinmed. 2005;  4 211-213
    PubMedSearch in Google Scholar
  • 15 Zouboulis CC, Seltmann H, Neitzel H, Orfanos CE. Establishment and characterization of an immortalized human sebaceous gland cell line (SZ95).  J Invest Dermatol. 1999;  113 1011-1020
    CrossrefPubMedSearch in Google Scholar
  • 16 Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham A, Fusenig NE. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.  J Cell Biol. 1988;  106 761-778
    CrossrefPubMedSearch in Google Scholar
  • 17 Zouboulis CC, Garbe C, Krasagakis K, Krüger S, Schröder K, Orfanos CE. A fluoremetric rapid microassay to identify anti-proliferative compounds for human melanoma cells in vitro.  Melanoma Res. 1991;  1 91-95
    PubMedSearch in Google Scholar
  • 18 Thiboutot D, Martin P, Volikos L, Gilliland K. Oxidative activity of the type 2 isozyme of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) predominates in human sebaceous glands.  J Invest Dermatol. 1998;  111 390-395
    CrossrefPubMedSearch in Google Scholar
  • 19 Rosenfield RL, Deplewski D, Kentsis A, Ciletti N. Mechanisms of androgen induction of sebocyte differentiation.  Dermatology. 1998;  196 43-46
    CrossrefPubMedSearch in Google Scholar
  • 20 Akamatsu H, Zouboulis CC, Orfanos CE. Control of human sebocyte proliferation in vitro by testosterone and 5-alpha-dihydrotestosterone is dependent on the localization of the sebaceous glands.  J Invest Dermatol. 1992;  99 509-511
    CrossrefPubMedSearch in Google Scholar
  • 21 Ellsworth K, Azzolina B, Baginsky W, Bull H, Chang B, Cimis G, Mitra S, Toney J, Bakshi RK, Rasmusson GR, Tolman RL, Harris GS. MK386: a potent, selective inhibitor of the human type 1 5alpha-reductase.  J Steroid Biochem Mol Biol. 1996;  58 377-384
    CrossrefPubMedSearch in Google Scholar
  • 22 Schwartz JI, Tanaka WK, Wang DZ, Ebel DL, Geissler LA, Dallob A, Hafkin B, Gertz BJ. MK-386, an inhibitor of 5alpha-reductase type 1, reduces dihydrotestosterone concentrations in serum and sebum without affecting dihydrotestosterone concentrations in semen.  J Clin Endocrinol Metab. 1997;  82 1373-1377
    CrossrefPubMedSearch in Google Scholar
  • 23 Tian G, Stuart JD, Moss ML, Domanico PL, Bramson HN, Patel IR, Kadwell SH, Overton LK, Kost TA, Mook Jr RA. et al . 17β-(N-tert-butylcarbomoyl)-4-aza-5α-androstane-1-en-3-one is an active site directed slow time-dependent inhibitor of human steroid 5α-reductase 1.  Biochemistry J. 1994;  33 2291-2296
    PubMedSearch in Google Scholar
  • 24 Imperato-Mc Ginley J, Gautier T, Cai L-Q, Yee B, Epstein J, Pochi P. The androgen control of sebum production. Studies of subjects with dihydrotestosterone deficiency and complete androgen insensitivity.  J Clin Endocrinol Metab. 1993;  76 524-528
    CrossrefPubMedSearch in Google Scholar
  • 25 Dallob AL, Sadick NS, Unger W, Lipert S, Geissler LA, Gregoire SL, Nguyen HH, Moore EC, Tanaka WK. The effect of finasteride a 5α-reductase inhibitor, on scalp skin testosterone and dihydrotestosterone concentration in patients with male pattern baldness.  J Clin Endocrinol Metab. 1994;  79 703-706
    CrossrefPubMedSearch in Google Scholar
  • 26 Kaufman KD, Olsen EA, Whiting D, Savin R, De Villez R, Bergfeld W, Price VH, Van Neste D, Roberts JL, Hordinsky M, Shapiro J, Binkowitz B, Gormley GJ. Finasteride in the treatment of men with androgenic alopecia. Finasteride male pattern hair loss study group.  J Am Acad Dermatol. 1998;  39 578-589
    CrossrefPubMedSearch in Google Scholar
  • 27 Wong IL, Morris RS, Chang L, Spahn MA, Stanczyk FZ, Lobo RA. A prospective randomized trial comparing finasteride to spironolactone in the treatment of hirsute woman.  J Clin Endocrinol Metab. 1995;  80 233-238
    CrossrefPubMedSearch in Google Scholar
  • 28 Di Salle E, Guidici D, Briatico G, Ornati G, Panzeri A. Hormonal effects of turosteride, a 5α-reductase inhibitor in the rat.  J Steroid Biochem Mol Biol. 1993;  46 549-555
    CrossrefPubMedSearch in Google Scholar
  • 29 Iehle C, Delso S, Guirou O, Tate R, Raynaud JP, Martin PM. Human prostatic steroid 5α-reductase isoforms - a comparative study of selective inhibitors.  J Steroid Biochem Molec Biol. 1995;  54 273-279
    PubMedSearch in Google Scholar
  • 30 Mowszowicz I, Wright F, Vincens M, Rigaud C, Nahoud K, Mavier P, Guillemant S, Kuttenn F, Mauvais-Jarvis P. Androgen metabolism in hirsute patients treated with cyproterone acetate.  J Steroid Biochem. 1984;  20 757-761
    CrossrefPubMedSearch in Google Scholar
  • 31 Corvol P, Michaud A, Menard J, Freifeld M, Mahoudeau J. Antiandrogenic affects of spironolactones: mechanism of action.  Endocrinology. 1975;  97 52-58
    PubMedSearch in Google Scholar
  • 32 Serafini PC, Catalino J, Lobo RA. The effect of spironolactone on genital skin 5α-reductase activity.  J Steroid Biochem. 1985;  23 191-194
    CrossrefPubMedSearch in Google Scholar
  • 33 Bouton MM, Lecaque D, Secchi J, Tournemine C. Effect of a new topically active antiandrogen (RU 38882) on the rat sebaceous gland: comparison with cyproterone acetate.  J Invest Dermatol. 1986;  86 163-167
    CrossrefPubMedSearch in Google Scholar
  • 34 Battmann T, Bonfils A, Branche C, Humbert J, Goubet F, Teutsch G, Philibert D. RU 58841, a new specific topical antiandrogen: a candidate of choice for the treatment of acne, androgenetic alopecia and hirsutism.  J Steroid Biochem Molec Biol. 1994;  48 55-60
    CrossrefPubMedSearch in Google Scholar
  • 35 Fimmel S, Saborowski A, Térouanne B, Sultan C, Zouboulis CC. Inhibition of the androgen receptor by antisense oligonucleotides regulates the biological activity of androgens in SZ95 sebocytes.  Horm Metab Res. 2007;  , in press
    PubMedSearch in Google Scholar
  • 36 Zouboulis CC, Eady A, Philpott M, Goldsmith LA, Orfanos C, Cunliffe WJ, Rosenfield R. What is the pathogenesis of acne?.  Exp Dermatol. 2005;  14 143-152
    CrossrefPubMedSearch in Google Scholar

1 The work was performed at the Department of Dermatology, University Medical Center Benjamin Franklin, Freie Universitaet Berlin, Berlin, Germany

Correspondence

C. C. Zouboulis

Departments of Dermatology and Immunology

Dessau Medical Center

Dessau

Germany

Phone: +49/340/501 40 00

Fax: +49/340/501 40 25

Email: christos.zouboulis@klinikum-dessau.de