The International Journal of Biochemistry & Cell Biology 41 (2009) 982–985

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The International Journal of Biochemistry

& Cell Biology
journal homepage: www.elsevier.com/locate/biocel

Molecules in focus

Vitamin D
Katie M. Dixon ∗ , Rebecca S. Mason
Department of Physiology and Bosch Institute, University of Sydney, NSW 2006 Australia

a r t i c l e i n f o a b s t r a c t

Article history: The primary source of vitamin D is the skin, following exposure to ultraviolet radiation. Vitamin D is
Received 3 June 2008 well known for its effects on stimulating calcium absorption and is thus essential for maintenance of
Received in revised form 10 June 2008 normal bone. It is also important for muscle function and has more recently been implicated in protection
Accepted 10 June 2008
against several diseases including diabetes. Different pathways of action have been described for vitamin
Available online 3 August 2008
D compounds and various analogs specific to these pathways have demonstrated potential for therapeutic
use. Recent studies suggest a novel role for vitamin D compounds in protection against cancer, a proposal
Keywords:
supported by substantial epidemiological evidence.
Vitamin D
1,25-Dihydroxyvitamin D3 © 2008 Elsevier Ltd. All rights reserved.
Cancer

1. Introduction 2. Structure

Following the discovery of vitamin D by Mellanby in 1920, its
physiological significance has been progressively realized. It was
originally classified as a “vitamin” essential for normal skeletal
development and maintenance of calcium homeostasis. However, it
has since been revealed that vitamin D is not strictly a vitamin since
it can be synthesized in the body as a result of UVR exposure. Vita-
min D is rather a steroid, more specifically, a secosteroid, indicating
that one of the rings of the cyclopentanoperhydrophenanthrene
ring structure (in the case of vitamin D, the 9–10 carbon–carbon
bond of ring B) is broken.
Vitamin D can be obtained from dietary sources or can be syn-
thesized in the body. Dietary vitamin D is available in two forms:
vitamin D2 (ergocalciferol) from plant sources and vitamin D3
(cholecalciferol) from animal sources, both of which are collectively
termed vitamin D. However, dietary sources generally account for
a very small amount of the total vitamin D in the body. Vitamin D in
the form of vitamin D3 can be made from 7-dehydrocholesterol (7-
DHC) in the skin by exposure to ultraviolet light, mainly from light
in the UVB spectrum (Holick, 2004). Vitamin D is a prohormone,
converted ultimately to the active form 1␣,25-dihydroxyvitamin
D3 (1,25(OH)2 D3 ).
∗ Corresponding author. Tel.: +61 2 9351 2561; fax: +61 2 9351 2510.
E-mail address: katied@physiol.usyd.edu.au (K.M. Dixon).
The molecular structure of vitamin D is closely allied to that
of classical steroid hormones. Vitamin D and all its metabolites,
including the steroid hormone 1,25(OH)2 D3 are, in comparison to
other steroid hormones, unusually conformationally flexible. This
arises from the flexible properties of the three major regions of
the molecule (Fig. 1A). The side chain has a 360◦ rotation around
each of the five carbon–carbon single bonds; the A-ring undergoes
a cyclohexane-like chair–chair interconversion, which changes the
orientation of the 1␣-hydroxyl and 3␤-hydroxyl groups; and the
broken B-ring has 360◦ rotation around the 6–7 single carbon bond.
This flexibility results in rapid conformational changes, occurring
millions of times per second. Since vitamin D can undergo rota-
tion about the 6–7 carbon–carbon bond, a wide variety of potential
ligand shapes can be generated, extending from the 6-s-cis steroid-
like conformation to the 6-s-trans extended steroid conformation
(Fig. 1B) (Norman et al., 2001).
3. Expression, activation and turnover
Upon exposure of the skin to ultraviolet light, the 9–10
carbon–carbon bond of 7-DHC is cleaved, forming pre-vitamin
D3 , which at body temperature thermally isomerizes over several
hours to days to vitamin D3 (Holick, 2004). However, the vitamin
D molecule itself has no demonstrated intrinsic biological activity.
Vitamin D3 binds to the vitamin D-binding protein, which selec-
tively transports vitamin D in the bloodstream to target cells of
the vitamin D endocrine system for metabolism. It then undergoes
two sequential biochemical alterations involving the addition of
two hydroxyl groups by cytochrome P450 enzymes. The first of

1357-2725/$ – see front matter © 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biocel.2008.06.016
 K.M. Dixon, R.S. Mason / The International Journal of Biochemistry & Cell Biology 41 (2009) 982–985
Fig. 1. Structure and conformational flexibility of 1,25(OH)2 D3 . (A) Structure and
sites of flexibility. (B) 6-s-cis steroid-like conformation and 6-s-trans extended
steroid conformation.
these hydroxylation reactions occurs in the liver, where vitamin
D is hydroxylated by the enzyme 25-hydroxylase at the carbon-25
position to form 25-hydroxyvitamin D (25(OH)D). This compound
is further hydroxylated in the kidney by 25-hydroxyvitamin D-1␣-
hydroxylase (1␣-hydroxylase) to the biologically active metabolite
1,25(OH)2 D (Holick, 2004), which can then generate biological
responses in over 30 target tissues by acting as a ligand for the
receptors that mediate pathways for vitamin D action (Norman
et al., 2001). Prolonged sunlight exposure cannot result in exces-
sive production of vitamin D3 . During extended sun exposure, both
previtamin D3 and the thermal isomerization product, vitamin D3 ,
absorb solar UVR and are converted to several “overirradiation
products” (Holick, 2004).
The two vitamin D activation steps have been shown to occur
in several other tissues. Activity of 25-hydroxylase has also been
observed in the intestine, adrenal gland, lung, kidney and bone,
whilst 1␣-hydroxylation of 25(OH)D3 to form 1,25(OH)2 D3 also
occurs in many tissues, including colon, breast, prostate, lung, acti-
vated macrophages, and parathyroid cells (Holick, 2004). Recent
studies in vitamin D metabolism have revealed that the complete
pathway of vitamin D synthesis, from 7-DHC to 1,25(OH)2 D3 , can
also occur in epidermal cells (Lehmann et al., 2001; Bikle et al.,
1986). This may be of significance in the photoprotection of skin
cells by 1,25(OH)2 D3 .
983
4. Biological function
Vitamin D compounds are best known for their role in stimu-
lating calcium absorption and thus contributing to optimal bone
mineralization and reduced fracture risk. The role of vitamin D in
calcium homeostasis is highlighted in genetic mouse models of tar-
geted deletion of either the 1,25(OH)2 D3 -synthesizing enzyme, 25
hydroxyvitamin D-1␣-hydroxylase [1␣(OH)ase or CYP27B1], or of
the nuclear receptor for 1,25(OH)2 D3 , the vitamin D receptor (VDR).
Even with heroic calcium supplementation, which almost abol-
ishes most of the phenotype, these mice have reduced osteoblast
numbers and bone volume (Goltzman et al., 2004). Furthermore,
vitamin D compounds play an important role in muscle func-
tion, and more recently, adequate vitamin D status has been
implicated in protection against a number of diseases including
diabetes, infection and a number of cancers (Peterlik and Cross,
2005).
The major hormonally active product of the vitamin D endocrine
system is 1,25(OH)2 D3 . The various biological responses produced
by 1,25(OH)2 D3 depend upon possibly two classes of vitamin D
receptors which, with their shape-sensitive and stereo-selective
ligand binding domains, determine the signal transduction path-
ways that become activated (Norman et al., 2001).
It is well established that 1,25(OH)2 D3 can produce genomic bio-
logical responses via signal transduction pathways that utilize a
nuclear receptor (VDR) for 1,25(OH)2 D3 to regulate gene transcrip-
tion. The genomic effects depend upon the interaction between
1,25(OH)2 D3 and the VDR followed by interaction of the activated
steroid receptor complex in the nucleus with vitamin D response
elements (VDREs) present in the promoter regions of the regulated
genes which are either to be activated or repressed (Norman et al.,
2001). The biological responses that occur as a result of the genomic
pathway are not immediate since they involve time-consuming
processes such as gene transcription.
A rapid-acting, non-genomic pathway for 1,25(OH)2 D3 action
has been proposed to generate a variety of biological responses
within seconds to minutes. The receptor involved in this pathway,
however, has not yet been definitively identified. The first report of
a rapid response-related membrane binding protein/receptor for
1,25(OH)2 D3 was in the basal lateral membrane of chick epithelial
cells and was linked to the rapid calcium absorption from the duo-
denum, a process known as transcaltachia (Nemere et al., 1984).
One group has presented evidence of a membrane-associated
rapid response steroid-binding protein (1,25D-MARRS), identical to
ERp57, as the receptor involved in rapid responses by 1,25(OH)2 D3
(Nemere et al., 2004). Recent reports implicate an alternate pocket
in the ligand binding domain of the classical vitamin D receptor
(VDR) in the rapid response (Mizwicki et al., 2004). The binding
of 1,25(OH)2 D3 to its proposed rapid-response receptor can stimu-
late various signaling pathways including protein kinase C, cAMP,
intracellular calcium and MAP kinase (Norman et al., 2001).
The conformational flexibility of 1,25(OH)2 D3 enables it to gen-
erate a wide variety of ligand shapes for available receptor(s) in
the vitamin D endocrine system. Studies of the X-ray structure of
the VDR have shown that the preferred ligand shape for this recep-
tor, when 1,25(OH)2 D3 is present as a ligand, is that represented
by a twisted 6-s-trans bowl. Structure-function studies involving
a variety of analogs of 1,25(OH)2 D3 locked in a particular confor-
mational shape suggest that the preferred shape for the receptor
linked to rapid responses is the 6-s-cis conformation (Norman et
al., 2001). Conformationally flexible analogs can, in principle, ini-
tiate either genomic or rapid responses. The known agonists and
antagonists have been used to dissect signal pathways in various
systems. Fig. 2 summarizes the pathways of action for vitamin D
and its analogs.
984 K.M. Dixon, R.S. Mason / The International Journal of Biochemistry & Cell Biology 41 (2009) 982–985

Fig. 2. Pathways of action for 1,25(OH)2 D3 and analogs. In principle, 1,25(OH)2 D3 and conformationally flexible analogs can activate either the genomic pathway or rapid
response pathway. Genomic responses are initiated following interaction between the vitamin D compound and the VDR followed by interaction of the activated steroid
receptor complex in the nucleus with vitamin D response elements in the promoter regions of the genes which are either to be activated or repressed. Analogs locked in
the 6-s-cis conformation such as 1,25-dihydroxylumisterol3 (JN) are full agonists for the rapid pathway and can only weakly bind the VDR. A membrane-associated rapid
response steroid-binding protein (1,25D-MARRS) has been proposed as the receptor for the rapid response pathway though recent reports implicate an alternate pocket in
the ligand-binding domain of the classical VDR in rapid responses. Binding of vitamin D compounds to the receptor for rapid responses triggers activation of other signal
transduction pathways including MAP kinase.

5. Possible medical applications
Recent studies suggest a role for vitamin D compounds in the
prevention of skin cancer. The active metabolite 1,25(OH)2 D3 has
been shown to inhibit ultraviolet radiation (UVR)-induced cell loss
when added to cultured human skin cells prior to and/or immedi-
ately after UVR (Wong et al., 2004). The improved cell survival in
1,25(OH)2 D3 -treated cells does not appear to come at the expense
of increased DNA damage in remaining cells. It has also been
shown that 1,25(OH)2 D3 has a dose-dependent inhibitory effect on
cyclobutane pyrimidine dimers (CPD) following UVR (Wong et al.,
2004; De Haes et al., 2005).
The protective effects of 1,25(OH)2 D3 against UVR-induced skin
damage have also been observed in vivo. Topical application or
intraperitoneal injection of 1,25(OH)2 D3 prior to UVR (Hanada et
al., 1995), or topical application immediately after UVR only (Gupta
et al., 2007) reduced apoptotic sunburn cells in mouse skin. Fur-
thermore, a reduction in skin CPD has been noted in mice treated
topically with 1,25(OH)2 D3 (Dixon et al., 2005).
While high concentrations of vitamin D have been shown to
be immunosuppressive, vitamin D deficiency also causes immuno-
suppression (Yang et al., 1993). In mice, topical 1,25(OH)2 D3
protected against UVR-induced systemic immunosuppression
(Dixon et al., 2005). Furthermore, it has been shown that
1,25(OH)2 D3 dose-dependently suppresses epidermal interleukin-
6 which usually increases after UVR exposure (De Haes et al.,
2003).
Several studies support a role for the rapid response
pathway in protection against UVR-induced skin damage by
1,25(OH)2 D3 . Two 6-s-cis locked low calcemic rapid-acting
agonists, 1,25-dihydroxylumisterol3 (JN) and 1␣,25-dihydroxy-7-
dehydrocholesterol (JM), entirely mimicked the photoprotective
effects of 1,25(OH)2 D3 in human skin cells (Wong et al., 2004).
Furthermore, these effects were completely abolished by a rapid-
acting antagonist whilst a genomic antagonist had no effect. Of
significance, the low calcemic compound JN was shown to be effec-
tive in an in vivo model, reducing UVR-induced sunburn cells, CPD
and immunosuppression (Dixon et al., 2005).
 K.M. Dixon, R.S. Mason / The International Journal of Biochemistry & Cell Biology 41 (2009) 982–985
These studies raise the possibility of the use of low calcemic
vitamin D compounds in skin cancer prevention. For exam-
ple, 1␣-hydroxymethyl-16-ene-24,24-difluoro-25-hydroxy-26,27-
bis-homovitamin D3 or QW-1624F2-2 (QW) is a transcriptionally
active analog that is approximately 80–100 times less calciuric than
1,25(OH)2 D3 , and does not cause cachexia (Posner et al., 2004).
The agent QW has high anti-proliferative and pro-differentiating
activity and has been shown to inhibit chemical-induced skin
tumorigenesis and tumour latency (Kensler et al., 2000). QW has
been fast tracked by the United States Food and Drug Administra-
tion (USFDA) for approval for clinical use in humans.
A randomized trial in postmenopausal women receiving vitamin
D and/or calcium supplementation showed a decreased all-cancer
risk with improved vitamin D status (Lappe et al., 2007). Fur-
thermore, VDR−/− mice were shown to be more susceptible to
chemical-induced skin carcinogenesis than their wild-type coun-
terparts (Zinser et al., 2002).
There is substantial epidemiological evidence correlating
decreased sunlight exposure and/or dietary vitamin D intake with
increased incidence of breast, prostate and colon cancers in humans
(Holick, 2004; Peterlik and Cross, 2005). A cohort study of skin and
non-skin cancer patients was consistent with a role for vitamin D
in decreasing the risk of several solid cancers, particularly stomach,
colorectal, liver and gallbladder, pancreas, lung, breast, prostate,
bladder and kidney. Incidence of second solid primary cancers after
skin melanoma was significantly lower for countries with higher
sunlight exposure than for those with lower sunlight exposure.
The difference was more obvious after non-melanoma skin can-
cers, which is consistent with earlier reports that non-melanoma
skin cancers reflect cumulative sun exposure, and thus effec-
tive vitamin D production. Conversely, melanoma is more related
to early intermittent exposure and sunburn (Tuohimaa et al.,
2007).
The studies outlined support the use of vitamin D compounds
in both therapeutic and preventative approaches to cancer. Vitamin
D synthesis and metabolism in skin may contribute to endogenous
photoprotection. Synthesis of vitamin D analogs with less calcemic
side effects is already resulting in the development of a novel class
of anticancer agents. Finally, the epidemiological data highlight the
importance of vitamin D status in protection from cancers.
Acknowledgements
This work was supported by the National Health and Medical
Research Council of Australia and the Cancer Council of New South
Wales. K.M. Dixon was the recipient of a Cancer Institute N.S.W.
Research Scholar Award. The authors apologize for the omission of
references due to space limitations.
References
Bikle DD, Nemanic MK, Gee E, Elias P. 1,25-Dihydroxyvitamin D3 production by
human keratinocytes. Kinetics and regulation. Journal of Clinical Investigation
1986;78:557–66.
985
De Haes P, Garmyn M, Degreef H, Vantieghem K, Bouillon R, Segaert S. 1,25-
Dihydroxyvitamin D3 inhibits ultraviolet B-induced apoptosis, Jun kinase
activation, and interleukin-6 production in primary human keratinocytes. Jour-
nal of Cellular Biochemistry 2003;89:663–73.
De Haes P, Garmyn M, Verstuyf A, De Clercq P, Vandewalle M, Degreef H, et al.
1,25-Dihydroxyvitamin D3 and analogues protect primary human keratinocytes
against UVB-induced DNA damage. Journal of Photochemistry & Photobiology B
Biology 2005;78:141–8.
Dixon KM, Deo SS, Wong G, Slater M, Norman AW, Bishop JE, et al. Skin cancer
prevention: a possible role of 1,25dihydroxyvitamin D3 and its analogs. Journal
of Steroid Biochemistry & Molecular Biology 2005;97:137–43.
Goltzman D, Miao D, Panda DK, Hendy GN. Effects of calcium and of the Vitamin
D system on skeletal and calcium homeostasis: lessons from genetic models.
Journal of Steroid Biochemistry & Molecular Biology 2004;89/90:485–9.
Gupta R, Dixon KM, Deo SS, J. Holliday C, Slater M, Halliday GM, et al. Photoprotection
by 1,25 dihydroxyvitamin D3 is associated with an increase in p53 and a decrease
in nitric oxide products. Journal of Investigative Dermatology 2007;127:
707–15.
Hanada K, Sawamura D, Nakano H, Hashimoto I. Possible role of 1,25-
dihydroxyvitamin D3 -induced metallothionein in photoprotection against UVB
injury in mouse skin and cultured rat keratinocytes. Journal of Dermatological
Science 1995;9:203–8.
Holick MF. Vitamin D: importance in the prevention of cancers, type 1 dia-
betes, heart disease, and osteoporosis. American Journal of Clinical Nutrition
2004;79:362–71 [erratum appears in Am J Clin Nutr 2004;79(May (5)):890].
Kensler TW, Dolan PM, Gange SJ, Lee JK, Wang Q, Posner GH. Conceptually new
deltanoids (vitamin D analogs) inhibit multistage skin tumorigenesis. Carcino-
genesis 2000;21:1341–5.
Lappe JM, Travers-Gustafson D, Davies KM, Recker RR, Heaney RP. Vitamin D and
calcium supplementation reduces cancer risk: results of a randomized trial.
American Journal of Clinical Nutrition 2007;85:1586–91.
Lehmann B, Genehr T, Knuschke P, Pietzsch J, Meurer M. UVB-induced conversion of
7-dehydrocholesterol to 1alpha,25-dihydroxyvitamin D3 in an in vitro human
skin equivalent model. Journal of Investigative Dermatology 2001;117:1179–85.
Mizwicki MT, Keidel D, Bula CM, Bishop JE, Zanello LP, Wurtz JM, et al. Identifica-
tion of an alternative ligand-binding pocket in the nuclear vitamin D receptor
and its functional importance in 1alpha, 25(OH)2 -vitamin D3 signaling. Pro-
ceedings of the National Academy of Sciences of the United States of America
2004;101:12876–81.
Nemere I, Yoshimoto Y, Norman AW, Nemere I, Yoshimoto Y, Norman AW. Cal-
cium transport in perfused duodena from normal chicks: enhancement within
fourteen minutes of exposure to 1,25-dihydroxyvitamin D3 . Endocrinology
1984;115:1476–83.
Nemere I, Farach-Carson MC, Rohe B, Sterling TM, Norman AW, Boyan BD, et al.
Ribozyme knockdown functionally links a 1,25(OH)2 D3 membrane binding pro-
tein (1,25D3 -MARRS) and phosphate uptake in intestinal cells. Proceedings of the
National Academy of Sciences of the United States of America 2004;101:7392–7.
Norman AW, Henry HL, Bishop JE, Song XD, Bula C, Okamura WH. Different shapes
of the steroid hormone 1alpha, 25(OH)(2)-vitamin D(3) act as agonists for two
different receptors in the vitamin D endocrine system to mediate genomic and
rapid responses. Steroids 2001;66:147–58.
Peterlik M, Cross HS. Vitamin D and calcium deficits predispose for multiple chronic
diseases. European Journal of Clinical Investigation 2005;35:290–304.
Posner GH, Jeon HB, Sarjeant A, Riccio ES, Doppalapudi RS, Kapetanovic IM, et
al. Low-calcemic, efficacious, 1␣,25-dihydroxyvitamin D3 analog QW-1624F2 -
2: calcemic dose–response determination, preclinical genotoxicity testing, and
revision of A-ring stereochemistry. Steroids 2004;69:757–62.
Tuohimaa P, Pukkala E, Scelo G, Olsen JH, Brewster DH, Hemminki K, et al. Does
solar exposure, as indicated by the non-melanoma skin cancers, protect from
solid cancers: vitamin D as a possible explanation. European Journal of Cancer
2007;43:1701–12.
Wong G, Gupta R, Dixon KM, Deo SS, Choong SM, Halliday GM, et al. 1,25-
Dihydroxyvitamin D and three low-calcemic analogs decrease UV-induced DNA
damage via the rapid response pathway. Journal of Steroid Biochemistry &
Molecular Biology 2004;89/90:567–70.
Yang S, Smith C, Prahl JM, Luo X, DeLuca HF. Vitamin D deficiency sup-
presses cell-mediated immunity in vivo. Archives of Biochemistry & Biophysics
1993;303:98–106.
Zinser GM, Sundberg JP, Welsh J. Vitamin D(3) receptor ablation sensitizes skin to
chemically induced tumorigenesis. Carcinogenesis 2002;23:2103–9.