• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. Apr 1990; 85(4): 1296–1303.
PMCID: PMC296566

1,25(OH)2 vitamin D3 stimulates membrane phosphoinositide turnover, activates protein kinase C, and increases cytosolic calcium in rat colonic epithelium.


The hormonal form of vitamin D, 1,25(OH)2 vitamin D3 [1,25(OH)2D3], regulates colonic calcium absorption and colonocyte proliferation and differentiation. In this study, we have examined the effect of 1,25(OH)2D3 on membrane phosphoinositide turnover, protein kinase C activation, and regulation of intracellular calcium concentration [( Ca+2]i) in isolated rat colonic epithelium. In a concentration-dependent manner, 1,25(OH)2D3 stimulated breakdown of membrane phosphoinositides within 15 s, generating diacylglycerol and inositol 1,4,5-triphosphate (IP3). 1,25(OH)2D3 rapidly activated colonic protein kinase C, with maximal translocation of activity from the cytosol to the membrane occurring within 1 min of exposure to the secosteroid. Studies performed in isolated colonocytes with the fluorescent dye fura-2 demonstrated that 10(-8) M 1,25(OH)2D3 caused a rapid rise in [Ca+2]i which then transiently decreased before rising to a new plateau value. When these experiments were performed in a calcium-free buffer, an increase in [Ca+2]i was observed, but both the transient and secondary rise were diminished in magnitude, suggesting that 1,25(OH)2D3 may stimulate both release of intracellular calcium stores and calcium influx. 1,25(OH)2D3 stimulated [3H]thymidine uptake in rat colonocytes, 4 h after an in vivo injection. These studies indicate that 1,25(OH)2D3 exerts a rapid influence on membrane phosphoinositide metabolism which may mediate certain of the secosteroid's effects on colonocyte calcium transport and proliferation.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Norman AW, Roth J, Orci L. The vitamin D endocrine system: steroid metabolism, hormone receptors, and biological response (calcium binding proteins). Endocr Rev. 1982 Fall;3(4):331–366. [PubMed]
  • Niendorf A, Arps H, Dietel M. Effect of 1,25-dihydroxyvitamin D3 on human cancer cells in vitro. J Steroid Biochem. 1987;27(4-6):825–828. [PubMed]
  • Koeffler HP. Vitamin D: myeloid differentiation and proliferation. Haematol Blood Transfus. 1985;29:409–417. [PubMed]
  • Brehier A, Thomasset M. Human colon cell line HT-29: characterisation of 1,25-dihydroxyvitamin D3 receptor and induction of differentiation by the hormone. J Steroid Biochem. 1988 Feb;29(2):265–270. [PubMed]
  • Wargovich MJ, Lointier PH. Calcium and vitamin D modulate mouse colon epithelial proliferation and growth characteristics of a human colon tumor cell line. Can J Physiol Pharmacol. 1987 Mar;65(3):472–477. [PubMed]
  • Birge SJ, Alpers DH. Stimulation of intestinal mucosal proliferation by vitamin D. Gastroenterology. 1973 May;64(5):977–982. [PubMed]
  • Hylander E, Ladefoged K, Jarnum S. The importance of the colon in calcium absorption following small-intestinal resection. Scand J Gastroenterol. 1980;15(1):55–60. [PubMed]
  • Garland CF, Garland FC. Do sunlight and vitamin D reduce the likelihood of colon cancer? Int J Epidemiol. 1980 Sep;9(3):227–231. [PubMed]
  • Garland C, Shekelle RB, Barrett-Connor E, Criqui MH, Rossof AH, Paul O. Dietary vitamin D and calcium and risk of colorectal cancer: a 19-year prospective study in men. Lancet. 1985 Feb 9;1(8424):307–309. [PubMed]
  • Minghetti PP, Norman AW. 1,25(OH)2-vitamin D3 receptors: gene regulation and genetic circuitry. FASEB J. 1988 Dec;2(15):3043–3053. [PubMed]
  • Ben Nasr L, Monet JD, Lucas PA. Rapid (10-minute) stimulation of rat duodenal alkaline phosphatase activity by 1,25-dihydroxyvitamin D3. Endocrinology. 1988 Oct;123(4):1778–1782. [PubMed]
  • Rasmussen H, Matsumoto T, Fontaine O, Goodman DB. Role of changes in membrane lipid structure in the action of 1,25-dihydroxyvitamin D3. Fed Proc. 1982 Jan;41(1):72–77. [PubMed]
  • Kurnik BR, Huskey M, Hagerty D, Hruska KA. Vitamin D metabolites stimulate phosphatidylcholine transfer to renal brush-border membranes. Biochim Biophys Acta. 1986 Jun 13;858(1):47–55. [PubMed]
  • Brasitus TA, Dudeja PK, Eby B, Lau K. Correction by 1-25-dihydroxycholecalciferol of the abnormal fluidity and lipid composition of enterocyte brush border membranes in vitamin D-deprived rats. J Biol Chem. 1986 Dec 15;261(35):16404–16409. [PubMed]
  • Craven PA, Pfanstiel J, DeRubertis FR. Role of activation of protein kinase C in the stimulation of colonic epithelial proliferation and reactive oxygen formation by bile acids. J Clin Invest. 1987 Feb;79(2):532–541. [PMC free article] [PubMed]
  • Craven PA, DeRubertis FR. Role of activation of protein kinase C in the stimulation of colonic epithelial proliferation by unsaturated fatty acids. Gastroenterology. 1988 Sep;95(3):676–685. [PubMed]
  • Craven PA, Pfanstiel J, DeRubertis FR. Role of reactive oxygen in bile salt stimulation of colonic epithelial proliferation. J Clin Invest. 1986 Mar;77(3):850–859. [PMC free article] [PubMed]
  • Roediger WE. Utilization of nutrients by isolated epithelial cells of the rat colon. Gastroenterology. 1982 Aug;83(2):424–429. [PubMed]
  • Kraft AS, Anderson WB, Cooper HL, Sando JJ. Decrease in cytosolic calcium/phospholipid-dependent protein kinase activity following phorbol ester treatment of EL4 thymoma cells. J Biol Chem. 1982 Nov 25;257(22):13193–13196. [PubMed]
  • FOLCH J, LEES M, SLOANE STANLEY GH. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed]
  • BARTLETT GR. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed]
  • Preiss J, Loomis CR, Bishop WR, Stein R, Niedel JE, Bell RM. Quantitative measurement of sn-1,2-diacylglycerols present in platelets, hepatocytes, and ras- and sis-transformed normal rat kidney cells. J Biol Chem. 1986 Jul 5;261(19):8597–8600. [PubMed]
  • MacDonald ML, Mack KF, Richardson CN, Glomset JA. Regulation of diacylglycerol kinase reaction in Swiss 3T3 cells. Increased phosphorylation of endogenous diacylglycerol and decreased phosphorylation of didecanoylglycerol in response to platelet-derived growth factor. J Biol Chem. 1988 Jan 25;263(3):1575–1583. [PubMed]
  • Wright TM, Rangan LA, Shin HS, Raben DM. Kinetic analysis of 1,2-diacylglycerol mass levels in cultured fibroblasts. Comparison of stimulation by alpha-thrombin and epidermal growth factor. J Biol Chem. 1988 Jul 5;263(19):9374–9380. [PubMed]
  • Muldoon LL, Rodland KD, Forsythe ML, Magun BE. Stimulation of phosphatidylinositol hydrolysis, diacylglycerol release, and gene expression in response to endothelin, a potent new agonist for fibroblasts and smooth muscle cells. J Biol Chem. 1989 May 25;264(15):8529–8536. [PubMed]
  • Downes CP, Mussat MC, Michell RH. The inositol trisphosphate phosphomonoesterase of the human erythrocyte membrane. Biochem J. 1982 Apr 1;203(1):169–177. [PMC free article] [PubMed]
  • van Dongen CJ, Zwiers H, Gispen WH. Microdetermination of phosphoinositides in a single extract. Anal Biochem. 1985 Jan;144(1):104–109. [PubMed]
  • Brasitus TA. Lipid dynamics and protein-lipid interactions in rat colonic epithelial cell basolateral membranes. Biochim Biophys Acta. 1983 Feb 9;728(1):20–30. [PubMed]
  • Sage SO, Rink TJ. The kinetics of changes in intracellular calcium concentration in fura-2-loaded human platelets. J Biol Chem. 1987 Dec 5;262(34):16364–16369. [PubMed]
  • Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed]
  • Verbin RS, Farber E. Effect of cycloheximide on the cell cycle of the crypts of the small intestine of the rat. J Cell Biol. 1967 Dec;35(3):649–658. [PMC free article] [PubMed]
  • BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. [PMC free article] [PubMed]
  • Norman AW, Ross FP. Vitamin D seco-steroids: unique molecules with both hormone and possible membranophilic properties. Life Sci. 1979 Feb 26;24(9):759–769. [PubMed]
  • Eisman JA, Fragonas JC, McMenemy ML. Rapid turnover of the 1,25-dihydroxyvitamin D3 receptor in human target cells. Endocrinology. 1988 Apr;122(4):1613–1621. [PubMed]
  • Chisholm JC, Kim S, Tashjian AH., Jr Modulation by 1,25-dihydroxycholecalciferol of the acute change in cytosolic free calcium induced by thyrotropin-releasing hormone in GH4C1 pituitary cells. J Clin Invest. 1988 Mar;81(3):661–668. [PMC free article] [PubMed]
  • Baran DT, Kelly AM. Lysophosphatidylinositol: a potential mediator of 1,25-dihydroxyvitamin D-induced increments in hepatocyte cytosolic calcium. Endocrinology. 1988 Mar;122(3):930–934. [PubMed]
  • Alkon DL, Rasmussen H. A spatial-temporal model of cell activation. Science. 1988 Feb 26;239(4843):998–1005. [PubMed]
  • Kuno M, Gardner P. Ion channels activated by inositol 1,4,5-trisphosphate in plasma membrane of human T-lymphocytes. Nature. 1987 Mar 19;326(6110):301–304. [PubMed]
  • Kikkawa U, Nishizuka Y. The role of protein kinase C in transmembrane signalling. Annu Rev Cell Biol. 1986;2:149–178. [PubMed]
  • Melloni E, Pontremoli S, Michetti M, Sacco O, Sparatore B, Horecker BL. The involvement of calpain in the activation of protein kinase C in neutrophils stimulated by phorbol myristic acid. J Biol Chem. 1986 Mar 25;261(9):4101–4105. [PubMed]
  • Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. [PubMed]
  • Weinstein IB. The origins of human cancer: molecular mechanisms of carcinogenesis and their implications for cancer prevention and treatment--twenty-seventh G.H.A. Clowes memorial award lecture. Cancer Res. 1988 Aug 1;48(15):4135–4143. [PubMed]
  • Ways DK, Dodd RC, Bennett TE, Gray TK, Earp HS. 1,25-Dihydroxyvitamin D3 enhances phorbol ester-stimulated differentiation and protein kinase C-dependent substrate phosphorylation activity in the U937 human monoblastoid cell. Endocrinology. 1987 Nov;121(5):1654–1661. [PubMed]
  • Mezzetti G, Bagnara GP, Monti MG, Pernecco Casolo L, Bonsi L, Brunelli MA. Phorbol esters, but not the hormonal form of vitamin D, induce changes in protein kinase C during differentiation of human histiocytic lymphoma cell line (U-937). Life Sci. 1987 May 25;40(21):2111–2117. [PubMed]
  • Sasaki K, Chida K, Hashiba H, Kamata N, Abe E, Suda T, Kuroki T. Enhancement by 1 alpha,25-dihydroxyvitamin D3 of chemically induced transformation of BALB 3T3 cells without induction of ornithine decarboxylase or activation of protein kinase C1. Cancer Res. 1986 Feb;46(2):604–610. [PubMed]
  • Berridge MJ, Irvine RF. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. [PubMed]
  • Baran DT, Milne ML. 1,25 Dihydroxyvitamin D increases hepatocyte cytosolic calcium levels. A potential regulator of vitamin D-25-hydroxylase. J Clin Invest. 1986 May;77(5):1622–1626. [PMC free article] [PubMed]
  • Sugimoto T, Ritter C, Ried I, Morrissey J, Slatopolsky E. Effect of 1,25-dihydroxyvitamin D3 on cytosolic calcium in dispersed parathyroid cells. Kidney Int. 1988 Apr;33(4):850–854. [PubMed]
  • Lieberherr M. Effects of vitamin D3 metabolites on cytosolic free calcium in confluent mouse osteoblasts. J Biol Chem. 1987 Sep 25;262(27):13168–13173. [PubMed]
  • Yoshimoto Y, Nemere I, Norman AW. Hypercalcemia inhibits the rapid stimulatory effect on calcium transport in perfused duodena from normal chicks mediated in vitro by 1,25-dihydroxyvitamin D3. Endocrinology. 1986 Jun;118(6):2300–2304. [PubMed]
  • de Boland AR, Boland RL. Rapid changes in skeletal muscle calcium uptake induced in vitro by 1,25-dihydroxyvitamin D3 are suppressed by calcium channel blockers. Endocrinology. 1987 May;120(5):1858–1864. [PubMed]
  • Nemere I, Szego CM. Early actions of parathyroid hormone and 1,25-dihydroxycholecalciferol on isolated epithelial cells from rat intestine: I. Limited lysosomal enzyme release and calcium uptake. Endocrinology. 1981 Apr;108(4):1450–1462. [PubMed]
  • Putkey JA, Spielvogel AM, Sauerheber RD, Dunlap CS, Norman AW. Vitamin D-mediated intestinal calcium transport. Effects of essential fatty acid deficiency and spin label studies of enterocyte membrane lipid fluidity. Biochim Biophys Acta. 1982 May 21;688(1):177–190. [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...