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首页> 外文期刊>Journal of Cellular Physiology >The effect of differentiation on 1,25 dihydroxyvitamin D-mediated gene expression in the enterocyte-like cell line, Caco-2.
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The effect of differentiation on 1,25 dihydroxyvitamin D-mediated gene expression in the enterocyte-like cell line, Caco-2.

机译：1日分化的影响,25岁dihydroxyvitamin D-mediated基因表达Caco-2 enterocyte-like细胞系。

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摘要

We examined 1,25 dihydroxyvitamin D (1,25(OH)(2)D(3))-induced expression of 25-hydroxyvitamin D(3) 24-hydroxylase (CYP24) and apical calcium channel (TRPV6) mRNA levels in 2-, 9-, and 15-day cultures Caco-2 cells that model proliferating, post-proliferative, and differentiated enterocytes. 1,25(OH)(2)D(3)-induced (10 nM, 8 h) CYP24 and TRPV6 mRNA levels were significantly greater in differentiated and post-proliferative than proliferating Caco-2 cells (>16X and >3X, respectively). Neither CYP24 mRNA half-life nor induction of a -298 bp rat CYP24 promoter-luciferase reporter construct (10 nM 1,25(OH)(2)D(3), 24 h) were different between proliferating and post-proliferating Caco-2 cells. We next tested whether the blunted response of natural genes to 1,25(OH)(2)D(3) in proliferating Caco-2 cells is due to altered chromatin remodeling. VDR and coactivator protein levels do not increase with differentiation but the level of the co-repressor Alien falls by 50% with differentiation. Over-expression of Alien reduced 1,25(OH)(2)D(3)-induced activity of a minimal VDRE containing promoter-luciferase construct by more than 60% in differentiated Caco-2 cells while siRNA knockdown of Alien in proliferating Caco-2 cells increased 1,25(OH)(2)D(3)-induced CYP24 mRNA level by 40%. These observations suggest that Alien is a regulator of VDR-mediated gene transcription in Caco-2 cells. In addition, we found that 1,25(OH)(2)D(3)-induced association of VDR with chromatin and with the CYP24 promoter was lower in proliferating cells. This suggests that decreased recruitment of VDR to vitamin D response elements also contributes to the blunted transcriptional responsiveness to 1,25(OH)(2)D(3) in proliferating Caco-2 cells.
机译：我们检查了1,25 dihydroxyvitamin D(1,25 (OH)(2)(3))全身的表达人体内25 -羟维生素D (3) 24-hydroxylase (CYP24)和顶端钙通道(TRPV6)的mRNA水平2 -9和15天文化Caco-2细胞模型增殖、post-proliferative和差异化的肠上皮细胞。1,25 (OH)全身的D (2) (3) (10 nM, 8 h) CYP24和TRPV6 mRNA水平明显提高分化和post-proliferative比细胞增殖Caco-2 (16 x和> > 3 x,分别)。感应的老鼠CYP24 -298个基点promoter-luciferase记者构造(10海里1,25 (OH) (2) (3), 24 h)之间是不同的增殖和post-proliferating Caco-2细胞。响应自然的基因1,25 (OH) (2) (3)细胞增殖Caco-2将改变染色质重塑。水平与分化,但不增加若外星人的水平下降了50%与分化。减少1,25 (OH)(2)(3)全身的活动的最小VDRE包含promoter-luciferase构建差异化的60%以上Caco-2细胞虽然siRNA可拆卸的外星人细胞增殖Caco-2增加1,25 (OH)(2)(3)全身CYP24 mRNA水平40%。这些观察表明,外星人是一个监管机构VDR-mediated基因的转录Caco-2细胞。D 1,25 (OH)(2)(3)全身VDR与协会染色质和CYP24启动子是较低的在增殖细胞。减少招聘的VDR维生素D响应元素也有助于减弱转录响应能力,1,25 (OH) (2) (3)在增殖Caco-2细胞。

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