Heimo Syvälä

Antiproliferative Action of Vitamin D

During the past few years, it has become apparent that vitamin D may play an important role in malignant transformation. Epidemiological studies suggest that low vitamin D serum concentration increases especially the risk of hormone-related cancers. Experimentally, vitamin D suppresses the proliferation of normal and malignant cells and induces differentiation and apoptosis. In the present review we discuss the mechanisms whereby vitamin D regulates cell proliferation and whether it could be used in prevention and treatment of hyperproliferative disorders like cancers.

25-Hydroxyvitamin D3 is an active hormone in human primary prostatic stromal cells

According to the present paradigm, 1α,25‐dihydroxyvitamin D3 [1α,25‐(OH)2D3] is a biologically active hormone; whereas 25‐hydroxyvitamin D3 (25OHD3) is regarded as a prohormone activated through the action of 25‐hydroxyvitamin D3 1α‐hydroxylase (1α‐ hydroxylase). Although the role of vitamin D3 in the regulation of growth and differentiation of prostatic epithelial cells has been well studied, its action and metabolism in prostatic stroma are still largely unknown. We investigated the effects of 25OHD3 and 1α,25‐(OH)2D3 on two human stromal primary cultures termed P29SN and P32S. In a cell proliferation assay, 25OHD3 was found at physiological concentrations of 100–250 nM to inhibit the growth of both primary cultures, whereas 1α,25‐(OH)2D3 at a pharmacological concentration of 10 nM exhibited the growth‐inhibitory effects on P29SN cells but not on P32S cells. Quantitative real‐time RT‐PCR analysis revealed that both 25OHD3 and 1α,25‐(OH)2D3 induced 25‐hydroxyvitamin D3 24‐ hydroxylase (24‐hydroxylase) mRNA in a dose‐ and time‐dependent manner. By inhibiting 1α‐ hydroxylase and/or 24‐hydroxylase enzyme activities, the induction of 24‐hydroxylase mRNA by 250 nM 25OHD3 was clearly enhanced, suggesting that 1α‐hydroxylation is not a prerequisite for the hormonal activity of 25OHD3. Altogether our results suggest that 25OHD3 at a high but physiological concentration acts as an active hormone with respect to vitamin D3 responsive gene regulation and suppression of cell proliferation.

The role of Vitamin D3 metabolism in prostate cancer

Vitamin D deficiency increases risk of prostate cancer. According to our recent results, the key Vitamin D hormone involved in the regulation of cell proliferation in prostate is 25(OH) Vitamin D3. It is mainly acting directly through the Vitamin D receptor (VDR), but partially also through its 1alpha-hydroxylation in the prostate. A deficiency of 25(OH) Vitamin D is common especially during the winter season in the Northern and Southern latitudes due to an insufficient sun exposure, but Vitamin D deficient diet may partially contribute to it. A lack of Vitamin D action may also be due to an altered metabolism or Vitamin D resistance. Vitamin D resistance might be brought up by several mechanisms: Firstly, an increased 24-hydroxylation may increase the inactivation of hormonal Vitamin D metabolites resulting in a Vitamin D resistance. This is obvious in the cancers in which an oncogenic amplification of 24-hydroxykase gene takes place, although an amplification of this gene in prostate cancer has not yet been described. During the aging, the activity of 24-hydroxylase increases, whereas 1alpha-hydroxylation decreases. Furthermore, it is possible that a high serum concentration of 25(OH)D3 could induce 24-hydroxylase expression in prostate. Secondly, Vitamin D receptor gene polymorphism or defects may result in a partial or complete Vitamin D resistance. Thirdly, an overexpression or hyperphosphorylation of retinoblastoma protein may result in an inefficient mitotic control by Vitamin D. Fourthly, endogenous steroids (reviewed by [D.M. Peehl, D. Feldman, Interaction of nuclear receptor ligands with the Vitamin D signaling pathway in prostate cancer, J. Steroid Biochem. Mol. Biol. (2004)]) and phytoestrogens may modulate the expression of Vitamin D metabolizing enzymes. In summary, the local metabolism of hormonal Vitamin D seems to play an important role in the development and progression of prostate cancer.

Statins and prostate cancer prevention: where we are now, and future directions

Statins are cholesterol-lowering drugs that are widely used to prevent and treat atherosclerotic cardiovascular disease. Recent research from both in vitro and in vivo studies suggests that there is an association between the use of statins and a reduction in the incidence of and mortality from prostate cancer. Several mechanisms of action that might bring about these beneficial effects of statins have been proposed, most of which include direct effects of statins on intracellular signaling. In this Review we discuss the current knowledge on the use of statins to prevent prostate cancer. We will also look at future directions for clinical research on this topic.

Vitamin D and prostate cancer.

Our recent epidemiological study (Ahonen et al., Cancer Causes Control 11(2000) (847-852)) suggests that vitamin D deficiency may increase the risk of initiation and progression of prostate cancer. The nested case-control study was based on a 13-year follow-up of about 19000 middle-aged men free of clinically verified prostate cancer. More than one-half of the serum samples had 25OH-vitamin D (25-VD) levels below 50 nmol/l, suggesting VD deficiency. Prostate cancer risk was highest among the group of younger men (40-51 years) with low serum 25-VD, whereas low serum 25-VD appeared not to increase the risk of prostate cancer in older men (>51 years). This suggests that VD has a protective role against prostate cancer only before the andropause, when serum androgen concentrations are higher. The lowest 25-VD concentrations in the younger men were associated with more aggressive prostate cancer. Furthermore, the high 25-VD levels delayed the appearance of clinically verified prostate cancer by 1.8 years. Since these results suggest that vitamin D has a protective role against prostate cancer, we tried to determine whether full spectrum lighting (FSL) during working hours could increase serum 25-VD concentrations. After 1-month exposure, there was no significant increase in the serum 25-VD level, although there was a bias towards slightly increasing values in the test group as opposed to decreasing values in controls. There was no significant change in the skin urocanic acid production. The possibility to use FSL in cancer prevention is discussed. In order to clarify the mechanism of VD action on cell proliferation and differentiation, we performed studies with the rat and human prostates as well prostate cancer cell lines. It is possible that 25-VD may have a direct role in the host anticancer defence activity, but the metabolism of vitamin D in the prostate may also play an important role in its action. We raised antibodies against human 1alpha-hydroxylase and 24-hydroxylase. Our preliminary results suggest that vitamin D is actively metabolised in the prostate. Vitamin D appears to upregulate androgen receptor expression, whereas androgens seem to upregulate vitamin D receptor (VDR). This may at least partially explain the androgen dependence of VD action. VD alone or administered with androgen causes a suppression of epithelial cell proliferation. VD can activate mitogen-activated kinases, erk-1 and erk-2, within minutes and p38 within hours. Also, auto/paracrine regulation might be involved, since keratinocyte growth factor (mRNA and protein) was clearly induced by VD. Based on these studies, a putative model for VD action on cell proliferation and differentiation is presented.

Interaction of Factors Related to the Metabolic Syndrome and Vitamin D on Risk of Prostate Cancer

Background: Factors related to the metabolic syndrome and low levels of vitamin D have been implicated as risk factors for prostate cancer. Insofar, no studies have assessed their joint effects on prostate cancer risk. Methods: We studied (a) the associations of vitamin D with the metabolic syndrome factors body mass index, systolic and diastolic blood pressure, and high-density lipoprotein cholesterol (HDL-C) and (b) the prostate cancer risk associated with these factors and especially their joint effects with vitamin D on risk of prostate cancer. We did a longitudinal nested case-control study on 132 prostate cancer cases and 456 matched controls from a cohort of 18,939 Finnish middle-aged men from the Helsinki Heart Study. The odds ratios (OR) of prostate cancer were assessed via conditional logistic regression analysis. Results: Apart from HDL-C, there was no linear association between the metabolic syndrome factors and vitamin D levels. In univariate analysis, men in the highest quartiles of body mass index (>28 kg/m2) and systolic blood pressure (>150 mmHg) showed a modest increase in risks of prostate cancer, with ORs of 1.37 (P = 0.16) and 1.53 (P = 0.05) when compared with the three lower quartiles, but low HDL-C entailed no prostate cancer risk. However, with all three factors present, the OR was 3.36 (P = 0.02), and jointly with low vitamin D (≤40 nmol/L), the OR was 8.03 (P = 0.005) compared with those with no metabolic syndrome factors and intermediate levels of vitamin D. There was an interaction between vitamin D and the metabolic syndrome factors so that a clustering of these factors entailed high risk of prostate cancer but only if vitamin D level was low (≤40 nmol/L). If it was at intermediate levels, the metabolic syndrome factors entailed no prostate cancer risk. Conclusions: We conclude that the prostate cancer risk associated with factors related to the metabolic syndrome is strongly conditioned by levels of vitamin D. (Cancer Epidemiol Biomarkers Prev 2007;16(2):302–7)

Vestibular dysfunction in vitamin D receptor mutant mice

The vitamin D endocrine system is essential for calcium and bone homeostasis. Vitamin D deficits are associated with muscle weakness and osteoporosis, whereas vitamin D supplementation may improve muscle function, body sway and frequency of falls, growth and mineral homeostasis of bones. The loss of muscle strength and mass, as well as deficits in bone formation, lead to poor balance. Poor balance is one of the main causes of falls, and may lead to dangerous injuries. Here we examine balance functions in vitamin D receptor deficient (VDR-/-) mice, an animal model of vitamin D-dependent rickets type II, and in 1alpha-hydroxylase deficient (1alpha-OHase-/-) mice, an animal model of pseudovitamin D-deficiency rickets. Recently developed methods (tilting box, rotating tube test), swim test, and modified accelerating rotarod protocol were used to examine whether the absence of functional VDR, or the lack of a key vitamin D-activating enzyme, could lead to mouse vestibular dysfunctions. Overall, VDR-/- mice, but not 1alpha-OHase-/- mice, showed shorter latency to fall from the rotarod, smaller fall angle in the tilting box test, and aberrant poor swimming. These data suggest that VDR deficiency in mice is associated with decreased balance function, and may be relevant to poorer balance/posture control in humans with low levels of vitamin D.

Localization of 1,25-dihydroxyvitamin D3 receptor (VDR) expression in human prostate

1,25-dihydroxyvitamin D3 (1,25(OH)2D3) has been found to have a variety of physiological functions, including effects on growth and differentiation in normal and malignant cells. The antiproliferative effects of 1,25(OH)2D3 are reported to be mediated through the genomic signaling pathway by binding to a specific high affinity receptor protein, the 1,25-dihydroxyvitamin D3 receptor (VDR). VDR has been localized in a variety of tissues, but little is known about VDR distribution in human prostate. In this study, we raised an antibody against a synthetic peptide corresponding to amino acids 10-24 of human vitamin D receptor. The sequence selected for immunization is identical in human, rat and mouse VDR. Based on this antibody, we developed an immunohistochemical method suitable for studying VDR expression in paraffin-embedded tissue. The immunohistochemical staining was verified using classical target organs for vitamin D (kidney, intestine, skin). With this method, we studied VDR localization on paraffin-embedded human prostatic tissue obtained from 8 patients undergoing radical prostatectomy for urinary bladder cancer and demonstrate VDR expression in the secretory epithelial and few stromal cells of human prostate. The nuclear staining in the secretory epithelial cells was concentrated near the nuclear membrane and in discrete foci in the nucleoplasm. This suggests that effects of 1,25-dihydroxyvitamin D3 are mediated through VDR in these cells. Moreover our result indicates that there are strong variations in VDR expression between prostatic samples.

EVIDENCE FOR ENHANCED UBIQUITIN-MEDIATED PROTEOLYSIS OF THE CHICKEN PROGESTERONE RECEPTOR BY PROGESTERONE

Genomic actions of progesterone are mediated via A and B isoforms of the progesterone receptor (PR). One major factor controlling PR level is progesterone causing negative autoregulation (down-regulation) of the receptor protein. In this work we studied the mechanism whereby progesterone exerts its effects on PR level in the chicken oviduct. We found that progesterone does not markedly regulate PR mRNA expression. Furthermore, we demonstrate here for the first time that PR is a target for ubiquitylation and that the proportion of ubiquitylated PR is increased by progesterone treatment. Our data suggest that ligand-induced down-regulation of PR involves enhanced degradation of receptor protein by ubiquitin-proteasome system in vivo.

Distribution of progesterone receptor in chicken: novel target organs for progesterone and estrogen action.

Expression of progesterone receptor (PR) in various organs of sexually immature chickens and after estrogen treatment was studied by immunohistochemical and Western blotting analyses. Constitutive PR expression was observed in the mesothelium and stroma of the esophagus, proventriculus, liver, spleen, pancreas, heart and lung. In the urogenital tract, PR was expressed in the mesothelial and stromal cells and smooth muscle of blood vessels. Estrogen treatment induced PR expression in the stroma and smooth muscle of the gall bladder and in the epithelium and stroma of the trachea. In the ovary of immature chickens PR was localized in the epithelium, stroma and smooth muscle and was induced in the granulosal cells by estrogen. In most tissues there was more PR-B than PR-A expression and this PR-B dominance remained after estrogen treatment. These results suggest that progesterone and estrogen may have physiological effects on many organs outside the genital tract not previously known as steroid-target tissues.