title: Steroidogenesis in the human skin: 21-hydroxylation in cultured keratinocytes.
abstract: We have evaluated the metabolism of radiolabeled progesterone (P) by the microsomal fraction isolated from HaCaT keratinocytes. P was widely metabolized to different compounds that included DOC (5-7% conversion) thus demonstrated 21-hydroxylase (21-OHase) activity, a key step in adrenal synthesis of gluco- and mineralocorticoids. However, RT-PCR amplification for the CYPc21 transcript of the corresponding gene showed no evidence for gene expression in HaCaT cells suggesting that the 21-OHase enzyme present in keratinocytes is different from that described in adrenal gland. Further characterization showed that whereas estradiol stimulated markedly P metabolism by HaCaT microsomes, with generation of new unidentified compounds, Lineweaver-Burk analysis of keratinocyte 21-OHase activity showed that the K(m) and V(max) were unaffected by estrogen. The apparent K(m) was 0.6 microM without estradiol and 0.7 microM with estradiol, while the respective V(max) values were 60 and 76 nmol/l/min. To conclude, we found extensive metabolism of P in human keratinocytes, we also provide the first demonstration of 21-OHase activity in this cell system and further showed that it is coded by a gene different from the adrenal CYPc21.
authors.affiliation: Centro de Investigaciones Endocrinologicas, Hospital de Niños Ricardo Gutiérrez, Buenos Aires, Argentina.
authors.forename: D
authors.initials: D
authors.lastname: Rogoff
authors.forename: C E
authors.initials: CE
authors.lastname: Gomez-Sanchez
authors.forename: M F
authors.initials: MF
authors.lastname: Foecking
authors.forename: J
authors.initials: J
authors.lastname: Wortsman
authors.forename: A
authors.initials: A
authors.lastname: Slominski
mesh_terms: D000311:Adrenal Glands / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D002855:Chromatography, Thin Layer; D003900:Desoxycorticosterone / Q000378:metabolism; D004967:Estrogens / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D007700:Kinetics; D008861:Microsomes / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011374:Progesterone / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000378:metabolism*; D013255:Steroid 21-Hydroxylase / Q000378:metabolism*
chemical_list: D004967:Estrogens; D012333:RNA, Messenger; D003900:Desoxycorticosterone; D011374:Progesterone; D013255:Steroid 21-Hydroxylase