Skip to main content

Advertisement

Log in

25-Hydroxyvitamin D3 is a natural chemopreventive agent against carcinogen induced precancerous lesions in mouse mammary gland organ culture

  • Preclinical Study
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Despite the role of vitamin D3 endocrine system in prevention of mammary gland transformation in animal models, use of 1,25(OH)2D3 in clinical settings is precluded due to its toxicity in vivo. Therefore much effort has been placed in developing relatively non-toxic vitamin D analogs. Recently, with the discovery of the expression of 25-hydroxy vitamin D3 1α-hydroxylase (CYP27B1) in multiple extrarenal organs, the functional role of prohormone, 25-hydroxyvitamin D3 [25(OH)D3], has been redefined. Since 25(OH)D3 does not cause hypercalcemia and maintains relative high concentration in serum, it is possible that the prohormone can be converted to active hormone in mammary epithelial cells to provide chemopreventive effects. In the present study, we evaluated its functional significance using mouse mammary organ culture (MMOC) system. We first showed that 25(OH)D3 1α-hydroxylase is extensively expressed in mammary ductal epithelial cells at both protein and mRNA levels, which is a prerequisite for 25(OH)D3 to function in an autocrine/paracrine manner. However, we also observed that clotrimazol (1α-hydroxylase inhibitor) enhanced 25(OH)D3 -induced CYP24 expression in breast cancer cells. In mammary glands derived from 1α-hydroxylase knockout mice, 25(OH)D3 treatment in organ culture significantly induced CYP24 expression, indicating a potential direct effect of 25(OH)D3. In MMOC, 100–250 nM 25(OH)D3 suppressed both ovarian hormone-dependent and -independent mammary precancerous lesions (induced by DMBA) by more than 50%, while the active hormone 1,25(OH)2D3 (positive control) at 100 nM suppressed alveolar lesions by more than 80%. The inactive vitamin D3 (negative control) at 100 nM suppressed alveolar lesions by only 20% (P > 0.05). We found that 25(OH)D3 inhibits DMBA-induced mammary alveolar lesions (MAL) in a stage-specific manner: 25(OH)D3 mainly inhibits the promotion stage of lesion formation. We conclude that 25(OH)D3 could serve as a non-toxic natural chemopreventive agent for further development for breast cancer prevention.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Berry DM, Meckling-Gill KA (1999) Vitamin D analogs, 20-Epi-22-oxa-24a,26a,27a,-trihomo-1alpha,25(OH)2-vitamin D3, 1,24(OH)2-22-ene-24-cyclopropyl-vitamin D3 and 1alpha,25(OH)2-lumisterol3 prime NB4 leukemia cells for monocytic differentiation via nongenomic signaling pathways, involving calcium and calpain. Endocrinology 140:4779–4788

    Article  PubMed  CAS  Google Scholar 

  2. Milliken EL, Zhang X, Flask C, Duerk JL, MacDonald PN, Keri RA (2005) EB1089, a vitamin D receptor agonist, reduces proliferation and decreases tumor growth rate in a mouse model of hormone-induced mammary cancer. Cancer Lett 229:205–215

    Article  PubMed  CAS  Google Scholar 

  3. Ryhanen S, Jaaskelainen T, Mahonen A, Maenpaa PH (2003) Inhibition of MG-63 cell cycle progression by synthetic vitamin D3 analogs mediated by p27, Cdk2, cyclin E, and the retinoblastoma protein. Biochem Pharmacol 66: 495–504

    Article  PubMed  CAS  Google Scholar 

  4. Spina CS, Ton L, Yao M, Maehr H, Wolfe MM, Uskokovic M, Adorini L, Holick MF (2007) Selective vitamin D receptor modulators and their effects on colorectal tumor growth. J Steroid Biochem Mol Biol 103:757–762

    Article  PubMed  CAS  Google Scholar 

  5. Mehta RG, Moriarty RM, Mehta RR, Penmasta R, Lazzaro G, Constantinou A, Guo L (1997) Prevention of preneoplastic mammary lesion development by a novel vitamin D analogue, 1alpha-hydroxyvitamin D5. J Natl Cancer Inst 89:212–218

    Article  PubMed  CAS  Google Scholar 

  6. Mehta RG, Hawthorne M, Uselding L, Albinescu D, Moriarty R, Christov K, Mehta RR (2000) Prevention of N-methyl-N-nitrosourea-induced mammary carcinogenesis in rats by 1alpha-hydroxyvitamin D5. J Natl Cancer Inst 92: 1836–1840

    Article  PubMed  CAS  Google Scholar 

  7. Lowe LC, Guy M, Mansi JL, Peckitt C, Bliss J, Wilson RG, Colston KW (2005) Plasma 25-hydroxy vitamin D concentrations, vitamin D receptor genotype and breast cancer risk in a UK Caucasian population. Eur J Cancer 41:1164–1169

    Article  PubMed  CAS  Google Scholar 

  8. Grant WB, Garland CF (2004) A critical review of studies on vitamin D in relation to colorectal cancer. Nutr Cancer 48:115–123

    Article  PubMed  CAS  Google Scholar 

  9. Lipkin M, Newmark HL (1999) Vitamin D, calcium and prevention of breast cancer: a review. J Am Coll Nutr 8:392S–397S

    Google Scholar 

  10. Ainsleigh HG (1993) Beneficial effects of sun exposure on cancer mortality. Prev Med 22:132–140

    Article  PubMed  CAS  Google Scholar 

  11. Garland FC, Garland CF, Gorham ED, Young JF (1990) Geographic variation in breast cancer mortality in the United States: a hypothesis involving exposure to solar radiation. Prev Med 19:614–622

    Article  PubMed  CAS  Google Scholar 

  12. Zinser GM, Welsh J (2004) Accelerated mammary gland development during pregnancy and delayed postlactational involution in vitamin D3 receptor null mice. Mol Endocrinol 18:2208–2223

    Article  PubMed  CAS  Google Scholar 

  13. Cross HS, Kallay E, Farhan H, Weiland T, Manhardt T (2003) Regulation of extrarenal vitamin D metabolism as a tool for colon and prostate cancer prevention. Recent Results Cancer Res 164:413–425

    PubMed  CAS  Google Scholar 

  14. Friedrich M, Rafi L, Mitschele T, Tilgen W, Schmidt W, Reichrath J (2003) Analysis of the vitamin D system in cervical carcinomas, breast cancer and ovarian cancer. Recent Results Cancer Res 164:239–246

    PubMed  CAS  Google Scholar 

  15. Cross HS, Kallay E, Lechner D, Gerdenitsch W, Adlercreutz H, Armbrecht HJ (2004) Phytoestrogens and vitamin D metabolism: a new concept for the prevention and therapy of colorectal, prostate, and mammary carcinomas. J Nutr 134:1207S–1212S

    PubMed  Google Scholar 

  16. Lou YR, Laaksi I, Syvala H, Blauer M, Tammela TL, Ylikomi T, Tuohimaa P (2004) 25-hydroxyvitamin D3 is an active hormone in human primary prostatic stromal cells. FASEB J 18:332–344

    PubMed  CAS  Google Scholar 

  17. Prosser DE, Jones G (2004) Enzymes involved in the activation and inactivation of vitamin D. Trends Biochem Sci 29:664–73

    Article  PubMed  CAS  Google Scholar 

  18. Fu GK, Lin D, Zhang MY, Bikle DD, Shackleton CH, Miller WL, Portale AA (1997) Cloning of human 25-hydroxyvitamin D-1 alpha-hydroxylase and mutations causing vitamin D-dependent rickets type 1. Mol Endocrinol 11:1961–1970

    Article  PubMed  CAS  Google Scholar 

  19. Mehta RG (2004) Stage-specific inhibition of mammary carcinogenesis by 1alpha-hydroxyvitamin D5. Eur J Cancer 40:2331–2337

    Article  PubMed  CAS  Google Scholar 

  20. Mehta RG, Hawthorne ME, Steele VE (1997) Induction and prevention of carcinogen-induced precancerous lesions in mouse mammary gland organ culture. Methods Cell Sci 19:19–24

    Article  Google Scholar 

  21. Dardenne O, Prud’homme J, Arabian A, Glorieux FH, St-Arnaud R (2001) Targeted inactivation of the 25-hydroxyvitamin D(3)-1(alpha)-hydroxylase gene (CYP27B1) creates an animal model of pseudovitamin D-deficiency rickets. Endocrinology 142:3135–3141

    Article  PubMed  CAS  Google Scholar 

  22. Peng X, Mehta R, Wang S, Chellappan S, Mehta RG (2006) Prohibitin is a novel target gene of vitamin D involved in its antiproliferative action in breast cancer cells. Cancer Res 66:7361–7369

    Article  PubMed  CAS  Google Scholar 

  23. de Kok JB, Roelofs RW, Giesendorf BA, Pennings JL, Waas ET, Feuth T, Swinkels DW, Span PN (2005) Normalization of gene expression measurements in tumor tissues: comparison of 13 endogenous control genes. Lab Invest 85:154–159

    PubMed  Google Scholar 

  24. Friedrich M, Diesing D, Cordes T, Fischer D, Becker S, Chen TC, Flanagan JN, Tangpricha V, Gherson I, Holick MF, Reichrath J (2006) Analysis of 25-hydroxyvitamin D3–1alpha-hydroxylase in normal and malignant breast tissue. Anticancer Res 26:2615–2620

    PubMed  CAS  Google Scholar 

  25. Colston KW, Lowe LC, Mansi JL, Campbell MJ (2006) Vitamin D status and breast cancer risk. Anticancer Res 26:2573–2580

    PubMed  CAS  Google Scholar 

  26. Flynn G, Chung I, Yu WD, Romano M, Modzelewski RA, Johnson CS, Trump DL (2006) Calcitriol (1,25-dihydroxycholecalciferol) selectively inhibits proliferation of freshly isolated tumor-derived endothelial cells and induces apoptosis. Oncology 70:447–457

    Article  PubMed  CAS  Google Scholar 

  27. Swami S, Krishnan AV, Feldman D (2000) 1alpha,25-Dihydroxyvitamin D3 down-regulates estrogen receptor abundance and suppresses estrogen actions in MCF-7 human breast cancer cells. Clin Cancer Res 6:3371–3379

    PubMed  CAS  Google Scholar 

  28. Colston KW, Hansen CM (2001) Mechanisms implicated in the growth regulatory effects of vitamin D in breast cancer. Endocr Relat Cancer 9: 45–59

    Article  Google Scholar 

  29. Welsh J (2007) Vitamin D and prevention of breast cancer. Acta Pharmacol Sin 28:1373–1382

    Article  PubMed  CAS  Google Scholar 

  30. Townsend K, Banwell CM, Guy M, Colston KW, Mansi JL, Stewart PM, Campbell MJ, Hewison M (2005) Autocrine metabolism of vitamin D in normal and malignant breast tissue. Clin Cancer Res 11:3579–3586

    Article  PubMed  CAS  Google Scholar 

  31. Segersten U, Holm PK, Bjorklund P, Hessman O, Nordgren H, Binderup L, Akerstrom G, Hellman P, Westin G (2005) 25-Hydroxyvitamin D3 1alpha-hydroxylase expression in breast cancer and use of non-1alpha-hydroxylated vitamin D analogue. Breast Cancer Res 7:R980–R986

    Article  PubMed  CAS  Google Scholar 

  32. Turunen MM, Dunlop TW, Carlberg C, Vaisanen S (2007) Selective use of multiple vitamin D response elements underlies the 1 alpha,25-dihydroxyvitamin D3-mediated negative regulation of the human CYP27B1 gene. Nucleic Acids Res 35:2734–2747

    Article  PubMed  CAS  Google Scholar 

  33. Murayama A, Kim MS, Yanagisawa J, Takeyama K, Kato S (2004) Transrepression by a liganded nuclear receptor via a bHLH activator through co-regulator switching. EMBO J 23:1598–1608

    Article  PubMed  CAS  Google Scholar 

  34. Kemmis CM, Salvador SM, Smith KM, Welsh J (2006) Human mammary epithelial cells express CYP27B1 and are growth inhibited by 25-hydroxyvitamin D-3, the major circulating form of vitamin D-3. J Nutr 136:887–892

    PubMed  CAS  Google Scholar 

  35. Bouillon R, Okamura WH, Norman AW (1995). Structure-function relationships in the vitamin D endocrine system. Endocr Rev 16:200–257

    Article  PubMed  CAS  Google Scholar 

  36. Skowronski RJ, Peehl DM, Feldman D (1995) Actions of vitamin D3, analogs on human prostate cancer cell lines: comparison with 1,25-dihydroxyvitamin D3. Endocrinology 136:20–26

    Article  PubMed  CAS  Google Scholar 

  37. Ritter CS, Armbrecht HJ, Slatopolsky E, Brown AJ. (2006) 25-Hydroxyvitamin D(3) suppresses PTH synthesis and secretion by bovine parathyroid cells. Kidney Int 70: 654–659

    Article  PubMed  CAS  Google Scholar 

  38. Masuda S, Jones G (2006). Promise of vitamin D analogues in the treatment of hyperproliferative conditions. Mol Cancer Ther 5:797–808

    Article  PubMed  CAS  Google Scholar 

  39. Vieth R (1999) Vitamin D supplementation, 25-hydroxyvitamin D concentrations, and safety. Am J Clin Nutr 69:842–856

    PubMed  CAS  Google Scholar 

  40. Nykjaer A, Fyfe JC, Kozyraki R, Leheste JR, Jacobsen C, Nielsen MS, Verroust PJ, Aminoff M, de la Chapelle A, Moestrup SK, Ray R, Gliemann J, Willnow TE, Christensen EI. (2001) Cubilin dysfunction causes abnormal metabolism of the steroid hormone 25(OH) vitamin D(3). Proc Natl Acad Sci USA 98:13895–13900

    Article  PubMed  CAS  Google Scholar 

  41. Nykjaer A, Dragun D, Walther D, Vorum H, Jacobsen C, Herz J, Melsen F, Christensen EI, Willnow TE. (1999) An endocytic pathway essential for renal uptake and activation of the steroid 25-(OH) vitamin D3. Cell 96:507–515

    Article  PubMed  CAS  Google Scholar 

  42. Rowling MJ, Kemmis CM, Taffany DA, Welsh J. (2006) Megalin-mediated endocytosis of vitamin D binding protein correlates with 25-hydroxycholecalciferol actions in human mammary cells. J Nutr 136:2754–2759

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by NIH Public Health Service Grant R03 CA121365-02 (XP) and R01 CA82316 (RGM). Support for engineering of the CYP27B1K0 mice was provided by a grant from shriners of North America to R. St-Arnaud.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Rajendra G. Mehta.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Peng, X., Hawthorne, M., Vaishnav, A. et al. 25-Hydroxyvitamin D3 is a natural chemopreventive agent against carcinogen induced precancerous lesions in mouse mammary gland organ culture. Breast Cancer Res Treat 113, 31–41 (2009). https://doi.org/10.1007/s10549-008-9900-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-008-9900-0

Keywords

Navigation