Prevention of Preneoplastic Mammary Lesion Development by a Novel Vitamin D Analogue, 1 -Hydroxyvitamin D5
Authors: R. G. Mehta, R. R. Mehta, G. Lazzaro, A. Constantinou, R. M. Moriarty, R. Penmasta, L. Guo
Journal: JNCI Journal of the National Cancer Institute (1997)

Background: The form of vitamin D (vitamin D 3 ) in fortified milk and the provitamin D produced by the body undergo metabolic activation to a biologically active form, 1α,25-dihydroxyvitamin D 3 [1α,25(OH) 2 D 3 ]. This compound can induce cell differentiation and can prevent proliferation of cancer cells. However, because 1α,25(OH) 2 D 3 is hypercalcemic (effective in increasing serum calcium level), it is not suitable for use in cancer prevention or cancer therapy trials. Purpose: We synthesized a vitamin D 5 series analogue, 1ahydroxy, 24-ethyl-cholecalciferol, or 1α-hydroxyvitamin D 5 [1α(OH)D 5 ], and evaluated its chemopreventive activity in carcinogen-treated mammary glands in organ culture experiments. Methods: The analogue 1α(OH)Dα was synthesized from sitosterol acetate and was characterized by nuclear magnetic resonance. Its purity was evaluated by high-pressure liquid chromatography. The calcemic activities of vitamin D 3 and D 5 analogues were determined in vitamin D-deficient Sprague-Dawley rats. Mammary glands of BALB/c mice were placed in organ culture and treated with the carcinogen 7,12-dimethylbenz[a]anthracene (DMBA) to induce preneoplastic lesions. Vitamin D analogues were added to the culture medium at four different concentrations, and formation of mammary lesions was evaluated. The effects of 1α(OH)D 5 and 1α,25(OH) 2 D 3 on the expression of vitamin D receptors (VDRs) and transforming growth factor-β 1 (TGF-β1) were studied by immunohistochemistry. Statistical significance was determined by the chi-squared test. All reported P values were two-sided. Results: 1α,25(OH) 2 D 3 was fourfold more calcemic than 1α(OH)D 5 at a dose of 0.042 µg/kg per day in rats. Both 1α,25(OH) 2 D 3 and 1α(OH)D 5 inhibited the development of DMBA-induced preneoplastic lesions in mouse mammary glands compared with untreated glands. The effect of the vitamin D 3 analogue was observed at a much lower concentration (0.01 µM). Treatment with 1α(OH)D 5 resulted in a dose-related (0.01- 10.0 µM) inhibition without any toxicity, whereas the vitamin D 3 analogue was highly potent but toxic at concentrations of 1.0 µM or higher. Normal mouse mammary glands poorly express VDR and TGF-β1; incubation with 1α(OH)D 5 or 1α,25(OH) 2 D 3 dramatically induced their expression. Conclusions: This is the first report showing the possibility of chemoprevention by a vitamin D 5 series compound. We conclude that 1α(OH)D 5 is less calcemic than 1α,25(OH) 2 D 3 . It is nontoxic at a wide range of concentrations, but it is potent in inhibiting the development of preneoplastic lesions in mammary glands in organ culture. In addition, we show for the first time the induction of TGF-β1 in normal mammary tissues by a chemopreventive agent. Implications: 1α(OH)D 5 is a good candidate for in vivo chemoprevention studies. It may mediate its action by inducing expression of VDR and of TGF-β1, as is seen in other systems.