Please cite this article in press as: D. Somjen, et al., Vitamin D less-calcemic analog modulates the expression of estrogen receptors, vitamin D receptor and 1-hydroxylase 25-hydroxy vitamin D in human thyroid cancer cell lines, J. Steroid Biochem. Mol. Biol. (2012), http://dx.doi.org/10.1016/j.jsbmb.2012.09.015 ARTICLE IN PRESS G Model SBMB-3846; No. of Pages 3 Journal of Steroid Biochemistry & Molecular Biology xxx (2012) xxx–xxx Contents lists available at SciVerse ScienceDirect Journal of Steroid Biochemistry and Molecular Biology jo u r n al hom epage: www.elsevier.com/locate/jsbmb Review Vitamin D less-calcemic analog modulates the expression of estrogen receptors, vitamin D receptor and 1-hydroxylase 25-hydroxy vitamin D in human thyroid cancer cell lines D. Somjen a,∗ , M. Grafi-Cohen a , G.H. Posner b , O. Sharon a , Z. Kraiem a , N. Stern a a Institute of Endocrinology, Metabolism and Hypertension, Tel-Aviv Sourasky Medical Centre and Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel b Department of Chemistry, The Johns Hopkins University, Baltimore, MD, USA a r t i c l e i n f o Article history: Received 14 June 2012 Received in revised form 10 September 2012 Accepted 12 September 2012 Keywords: Thyroid cancer cell lines ERs VDR 1OHase JKF a b s t r a c t Estrogen receptors (ERs) are expressed in various “non-reproductive” cancer cell types. Some cancer types express 1-hydroxylase 25-hydroxy vitamin D (1OHase) whose product, 1,25(OH) 2 D 3 can retard cancer cell proliferation. Thyroid carcinoma cell growth is apparently promoted by estrogens, but whether or not this interaction is modified by vitamin D metabolites/analogs is presently unknown. Here we assessed the effect of a less calcemic vitamin D analog [JK 1624 F 2 -2 (JKF)] in three human thyroid cancer cell lines: ARO (anaplastic carcinoma), NPA (papillary carcinoma) and MRO (follicular carcinoma). (1) All cell lines expressed both ER and ER, vitamin D receptor (VDR) and 1OHase mRNA quantified by Real Time PCR. There was a general abundance of ER over ER expression, such that the ratio of ER to ER mRNA was >1000:1, 228:1 and 7.7:1 in ARO, MRO and NPA cells, respectively. (2) JKF up regulated ER expression in ARO (by 110 ± 15%) and MRO (by 280 ± 10%) but down regulated ER in NPA cells (by 40 ± 15%). The expression of VDR was up regulated by JKF in NPA (21 ± 6%), down regulated in ARO (-24 ± 7%) and not affected in MRO. (3) All three human thyroid cancer cell lines were found to express 1OHase, which was up regulated by JKF in MRO (350 ± 25%) and NPA (35 ± 8%) but down regulated in ARO (-20 ± 5%). This is the first report to describe direct regulation of VDR and 1OHase expression by a vitamin D analog in human thyroid cancer cells. A functional role for the vitamin D system in human thyroid cancer is suggested by the finding that the vitamin D analog can affect ERs expression which is in turn involved in estrogen-induced cell growth in an ER-type specific manner in these cells. © 2012 Elsevier Ltd. All rights reserved. Contents 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 2. Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 2.1. Statistical analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 3. Results .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 3.1. Expression of ER, ER, VDR and 1OHase in thyroid cancer cell lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 3.2. Effect of JKF on the expression of ER, ER, VDR and 1OHase in thyroid cancer cell lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 4. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 1. Introduction Several studies have indicated that vitamin D metabolites and analogs modulate the expression and activity of estrogen receptors ∗ Corresponding author at: Institute of Endocrinology, Metabolism and Hyper- tension, Tel.:-Aviv Sourasky Medical Centre, Tel-Aviv 64239, Israel. Tel.: +972 36973812; fax: +972 36973421. E-mail address: dalias@tasmc.health.gov.il (D. Somjen). (ERs), vitamin D receptor (VDR) and 1 hydroxy vitamin D hydrox- ylase (1OHase) in various normal and cancerous cells [1,2]. To date, such effects of the vitamin D less-calcemic analog JKF [3–5] were not analyzed in human thyroid cancer cell lines. Because thyroid tissue endogenous 1,25(OH) 2 D 3 , VDR expression and ER subtype expression are likely involved in thyroid cell/thyroid cancer cell growth, the effects of vitamin D analogs on these genes is of obvi- ous interest. Here we tested the expressions of ER, ER, VDR and 1OHase mRNA by Real Time PCR [2] in three human thyroid cancer 0960-0760/$ – see front matter © 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.jsbmb.2012.09.015