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. 1986 Aug;78(2):557–566. doi: 10.1172/JCI112609

1,25-Dihydroxyvitamin D3 production by human keratinocytes. Kinetics and regulation.

D D Bikle, M K Nemanic, E Gee, P Elias
PMCID: PMC423594  PMID: 2426308

Abstract

Human foreskin keratinocytes in vitro metabolize 25-hydroxyvitamin D3 to a number of metabolites, including 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). This metabolite remains mostly within the cell and does not accumulate in the medium under the conditions of these experiments. With time, 1,25(OH)2D3 is catabolized, and more polar metabolites appear in both the cells and the medium. The production of 1,25(OH)2D3 has an apparent Michaelis constant (Km) for 25-hydroxyvitamin D3 of 5.4 X 10(-8) M. The levels of 1,25(OH)2D3 within the cell are increased both by increased production and decreased catabolism when parathyroid hormone(1-34) and isobutylmethylxanthine are added. Exogenously added 1,25(OH)2D3 at concentrations as low as 10(-12) M reduces endogenous 1,25(OH)2D3 production, increases 1,25(OH)2D3 catabolism, and increases 24,25-dihydroxyvitamin D3 production by an actinomycin D-sensitive process. These data indicate that the regulation of 1,25(OH)2D3 production by keratinocytes is similar to, but not identical to the regulation of 1,25(OH)2D3 by the kidney.

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Selected References

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  1. Adams J. S., Sharma O. P., Gacad M. A., Singer F. R. Metabolism of 25-hydroxyvitamin D3 by cultured pulmonary alveolar macrophages in sarcoidosis. J Clin Invest. 1983 Nov;72(5):1856–1860. doi: 10.1172/JCI111147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Barbour G. L., Coburn J. W., Slatopolsky E., Norman A. W., Horst R. L. Hypercalcemia in an anephric patient with sarcoidosis: evidence for extrarenal generation of 1,25-dihydroxyvitamin D. N Engl J Med. 1981 Aug 20;305(8):440–443. doi: 10.1056/NEJM198108203050807. [DOI] [PubMed] [Google Scholar]
  4. Bikle D. D., Gee E., Halloran B., Haddad J. G. Free 1,25-dihydroxyvitamin D levels in serum from normal subjects, pregnant subjects, and subjects with liver disease. J Clin Invest. 1984 Dec;74(6):1966–1971. doi: 10.1172/JCI111617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bikle D. D., Nemanic M. K., Whitney J. O., Elias P. W. Neonatal human foreskin keratinocytes produce 1,25-dihydroxyvitamin D3. Biochemistry. 1986 Apr 8;25(7):1545–1548. doi: 10.1021/bi00355a013. [DOI] [PubMed] [Google Scholar]
  6. Bikle D. D., Rasmussen H. A comparison of the metabolism of 25-hydroxyvitamin D3 by chick renal tubules, homogenates and mitochondria. Biochim Biophys Acta. 1974 Oct 8;362(3):439–447. doi: 10.1016/0304-4165(74)90139-1. [DOI] [PubMed] [Google Scholar]
  7. Bikle D. D., Rasmussen H. The ionic control of 1,25-dihydroxyvitamin D3 production in isolated chick renal tubules. J Clin Invest. 1975 Feb;55(2):292–298. doi: 10.1172/JCI107932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bikle D. D., Rasmussen H. The metabolism of 25-hydroxycholecalciferol by isolated renal tubules in vitro as studied by a new chromatographic technique. Biochim Biophys Acta. 1974 Oct 8;362(3):425–438. doi: 10.1016/0304-4165(74)90138-x. [DOI] [PubMed] [Google Scholar]
  9. Brumbaugh P. F., Haussler D. H., Bursac K. M., Haussler M. R. Filter assay for 1alpha, 25-dihydroxyvitamin D3. Utilization of the hormone's target tissue chromatin receptor. Biochemistry. 1974 Sep 24;13(20):4091–4097. doi: 10.1021/bi00717a005. [DOI] [PubMed] [Google Scholar]
  10. Clemens T. L., Adams J. S., Horiuchi N., Gilchrest B. A., Cho H., Tsuchiya Y., Matsuo N., Suda T., Holick M. F. Interaction of 1,25-dihydroxyvitamin-D3 with keratinocytes and fibroblasts from skin of normal subjects and a subject with vitamin-D-dependent rickets, type II: a model for study of the mode of action of 1,25-dihydroxyvitamin D3. J Clin Endocrinol Metab. 1983 Apr;56(4):824–830. doi: 10.1210/jcem-56-4-824. [DOI] [PubMed] [Google Scholar]
  11. Esvelt R. P., DeLuca H. F., Wichmann J. K., Yoshizawa S., Zurcher J., Sar M., Stumpf W. E. 1,25-Dihydroxyvitamin D3 stimulated increase of 7,8-didehydrocholesterol levels in rat skin. Biochemistry. 1980 Dec 23;19(26):6158–6161. doi: 10.1021/bi00567a032. [DOI] [PubMed] [Google Scholar]
  12. Feldman D., Chen T., Cone C., Hirst M., Shani S., Benderli A., Hochberg Z. Vitamin D resistant rickets with alopecia: cultured skin fibroblasts exhibit defective cytoplasmic receptors and unresponsiveness to 1,25(OH)2D3. J Clin Endocrinol Metab. 1982 Nov;55(5):1020–1022. doi: 10.1210/jcem-55-5-1020. [DOI] [PubMed] [Google Scholar]
  13. Feldman D., Chen T., Hirst M., Colston K., Karasek M., Cone C. Demonstration of 1,25-dihydroxyvitamin D3 receptors in human skin biopsies. J Clin Endocrinol Metab. 1980 Dec;51(6):1463–1465. doi: 10.1210/jcem-51-6-1463. [DOI] [PubMed] [Google Scholar]
  14. Frankel T. L., Mason R. S., Hersey P., Murray E., Posen S. The synthesis of vitamin D metabolites by human melanoma cells. J Clin Endocrinol Metab. 1983 Sep;57(3):627–631. doi: 10.1210/jcem-57-3-627. [DOI] [PubMed] [Google Scholar]
  15. Fukase M., Birge S. J., Jr, Rifas L., Avioli L. V., Chase L. R. Regulation of 25 hydroxyvitamin D3 1-hydroxylase in serum-free monolayer culture of mouse kidney. Endocrinology. 1982 Mar;110(3):1073–1075. doi: 10.1210/endo-110-3-1073. [DOI] [PubMed] [Google Scholar]
  16. Gray T. K., Lester G. E., Lorenc R. S. Evidence for extra-renal 1 alpha-hydroxylation of 25-hydroxyvitamin D3 in pregnancy. Science. 1979 Jun 22;204(4399):1311–1313. doi: 10.1126/science.451538. [DOI] [PubMed] [Google Scholar]
  17. Henry H. L. The role of parathyroid hormone in the regulation of the metabolism of 25-hydroxyvitamin D3. Miner Electrolyte Metab. 1982 Sep-Oct;8(3-4):179–187. [PubMed] [Google Scholar]
  18. Holick M. F. The cutaneous photosynthesis of previtamin D3: a unique photoendocrine system. J Invest Dermatol. 1981 Jul;77(1):51–58. doi: 10.1111/1523-1747.ep12479237. [DOI] [PubMed] [Google Scholar]
  19. Hosomi J., Hosoi J., Abe E., Suda T., Kuroki T. Regulation of terminal differentiation of cultured mouse epidermal cells by 1 alpha,25-dihydroxyvitamin D3. Endocrinology. 1983 Dec;113(6):1950–1957. doi: 10.1210/endo-113-6-1950. [DOI] [PubMed] [Google Scholar]
  20. Howard G. A., Turner R. T., Sherrard D. J., Baylink D. J. Human bone cells in culture metabolize 25-hydroxyvitamin D3 to 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3. J Biol Chem. 1981 Aug 10;256(15):7738–7740. [PubMed] [Google Scholar]
  21. Labarca C., Paigen K. A simple, rapid, and sensitive DNA assay procedure. Anal Biochem. 1980 Mar 1;102(2):344–352. doi: 10.1016/0003-2697(80)90165-7. [DOI] [PubMed] [Google Scholar]
  22. Lambert P. W., Stern P. H., Avioli R. C., Brackett N. C., Turner R. T., Greene A., Fu I. Y., Bell N. H. Evidence for extrarenal production of 1 alpha ,25-dihydroxyvitamin D in man. J Clin Invest. 1982 Mar;69(3):722–725. doi: 10.1172/JCI110501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Littledike E. T., Horst R. L. Metabolism of vitamin D3 in nephrectomized pigs given pharmacological amounts of vitamin D3. Endocrinology. 1982 Dec;111(6):2008–2013. doi: 10.1210/endo-111-6-2008. [DOI] [PubMed] [Google Scholar]
  24. Mason R. S., Frankel T., Chan Y. L., Lissner D., Posen S. Vitamin D conversion by sarcoid lymph node homogenate. Ann Intern Med. 1984 Jan;100(1):59–61. doi: 10.7326/0003-4819-100-1-59. [DOI] [PubMed] [Google Scholar]
  25. Nemanic M. K., Whitney J., Arnaud S., Herbert S., Elias P. M. Vitamin D3 production by cultured human keratinocytes and fibroblasts. Biochem Biophys Res Commun. 1983 Sep 15;115(2):444–450. doi: 10.1016/s0006-291x(83)80164-8. [DOI] [PubMed] [Google Scholar]
  26. Puzas J. E., Turner R. T., Howard G. A., Baylink D. J. Cells isolated from embryonic intestine synthesize 1,25-dihydroxyvitamin-D3 and 24,25-dihydroxyvitamin-D3 in culture. Endocrinology. 1983 Jan;112(1):378–380. doi: 10.1210/endo-112-1-378. [DOI] [PubMed] [Google Scholar]
  27. Rasmussen H., Wong M., Bikle D., Goodman D. B. Hormonal control of the renal conversion of 25-hydroxycholecalciferol to 1,25-dihydroxycholecalciferol. J Clin Invest. 1972 Sep;51(9):2502–2504. doi: 10.1172/JCI107065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reeve L., Tanaka Y., DeLuca H. F. Studies on the site of 1,25-dihydroxyvitamin D3 synthesis in vivo. J Biol Chem. 1983 Mar 25;258(6):3615–3617. [PubMed] [Google Scholar]
  29. Rheinwald J. G., Green H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell. 1975 Nov;6(3):331–343. doi: 10.1016/s0092-8674(75)80001-8. [DOI] [PubMed] [Google Scholar]
  30. Rost C. R., Bikle D. D., Kaplan R. A. In vitro stimulation of 25-hydroxycholecalciferol 1 alpha-hydroxylation by parathyroid hormone in chick kidney slices: evidence for a role for adenosine 3',5'-monophosphate. Endocrinology. 1981 Mar;108(3):1002–1006. doi: 10.1210/endo-108-3-1002. [DOI] [PubMed] [Google Scholar]
  31. Shultz T. D., Fox J., Heath H., 3rd, Kumar R. Do tissues other than the kidney produce 1,25-dihydroxyvitamin D3 in vivo? A reexamination. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1746–1750. doi: 10.1073/pnas.80.6.1746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Spanos E., Barrett D. I., Chong K. T., MacIntyre I. Effect of oestrogen and 1,25-dihydroxycholecalciferol on 25-hydroxycholecalciferol metabolism in primary chick kidney-cell cultures. Biochem J. 1978 Jul 15;174(1):231–236. doi: 10.1042/bj1740231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Trechsel U., Bonjour J. P., Fleisch H. Regulation of the metabolism of 25-hydroxyvitamin D3 in primary cultures of chick kidney cells. J Clin Invest. 1979 Jul;64(1):206–217. doi: 10.1172/JCI109441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Turner R. T., Puzas J. E., Forte M. D., Lester G. E., Gray T. K., Howard G. A., Baylink D. J. In vitro synthesis of 1 alpha,25-dihydroxycholecalciferol and 24,25-dihydroxycholecalciferol by isolated calvarial cells. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5720–5724. doi: 10.1073/pnas.77.10.5720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Weisman Y., Harell A., Edelstein S., David M., Spirer Z., Golander A. 1 alpha, 25-Dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 in vitro synthesis by human decidua and placenta. Nature. 1979 Sep 27;281(5729):317–319. doi: 10.1038/281317a0. [DOI] [PubMed] [Google Scholar]
  36. Whitsett J. A., Ho M., Tsang R. C., Norman E. J., Adams K. G. Synthesis of 1,25-dihydroxyvitamin D3 by human placenta in vitro. J Clin Endocrinol Metab. 1981 Sep;53(3):484–488. doi: 10.1210/jcem-53-3-484. [DOI] [PubMed] [Google Scholar]

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