Effects of MAPK signaling on 1,25-dihydroxyvitamin D-mediated CYP24 gene expression in the enterocyte-like cell line, Caco-2.
Effects of MAPK signaling on 1,25-dihydroxyvitamin D-mediated CYP24 gene expression in the enterocyte-like cell line, Caco-2.
复制标题
DOI：
10.1002/jcp.21657
复制
发表时间：
2009-04
影响因子：
5.6
通讯作者：
Fleet, James C.
中科院分区：
文献类型：
--
作者：
Cui, Min;Zhao, Yan;Hance, Kenneth W.;Shao, Andrew;Wood, Richard J.;Fleet, James C.
我们研究了细胞外信号调节激酶（ERK）在1，25（OH）2D 3诱导分化的Caco-2细胞基因表达中的作用。1，25（OH）2D 3调节的25-羟基维生素D，24羟化酶（CYP 24）基因表达（天然基因和启动子构建体）通过改变ERK活性来强烈调节（即通过MEK抑制剂和显性负性（dn）ERK 1和ERK 2降低，由表皮生长因子激活），但ERK抑制对1，25（OH）2D 3调节的瞬时受体电位阳离子通道，亚家族V，成员6（TRPV 6）的表达。在增殖但未分化的Caco-2细胞中，由于分化细胞中ERK 5和Ets-1水平（总水平和磷酸化蛋白水平）降低，ERK 5介导的转录因子Ets-1磷酸化增强了1，25（OH）2D 3介导的CYP 24基因转录。MEK抑制降低了1，25（OH）2D 3诱导的3X-VDRE启动子活性，但对维生素D受体（VDR）与染色质的相关性没有影响，这表明辅激活剂募集在维生素D调节的CYP 24基因激活的ERK调节中发挥作用。染色质免疫沉淀分析显示，ERK 1/2靶点，介体1（MED 1），在1，25（OH）2D 3处理后被募集到CYP 24启动子，而不是TRPV 6启动子。MED 1磷酸化对ERK 1/2信号传导的激活剂和抑制剂敏感，并且MED 1 siRNA降低1，25（OH）2D 3调节的人CYP 24启动子活性。这表明ERK 1/2信号通过MED 1介导的事件增强了1，25（OH）2D 3对CYP 24启动子的作用。我们的数据表明，在肠细胞样Caco-2细胞中，ERK信号通路对1，25（OH）2D 3介导的基因诱导具有启动子特异性和细胞阶段特异性作用。
We examined the role of the Extracellular signal Regulated Kinases (ERK) in 1,25 dihydroxyvitamin D (1,25(OH)2D3)-induced gene expression in the differentiated Caco-2 cells. 1,25(OH)2D3-regulated expression of the 25-hydroxyvitamin D, 24 hydroxylase (CYP24) gene (both natural gene and promoter construct) was strongly modulated by altering ERK activity (i.e. reduced by MEK inhibitors and dominant negative (dn) ERK1 and ERK2, activated by Epidermal Growth Factor) but ERK inhibition had no effect on 1,25(OH)2D3-regulated expression of the transient receptor potential cation channel, subfamily V, member 6 (TRPV6). ERK5-mediated phosphorylation of the transcription factor Ets-1 enhanced 1,25(OH)2D3-mediated CYP24 gene transcription in proliferating but not differentiated Caco-2 cells due to reduced levels of ERK5 and Ets-1 (total and phospho-protein levels) in differentiated cells. MEK inhibition reduced 1,25(OH)2D3-induced 3X-VDRE promoter activity but had no impact on the association of vitamin D receptor (VDR) with chromatin suggesting a role for co-activator recruitment in ERK-modulation of vitamin D-regulated CYP24 gene activation. Chromatin immunoprecipitation assays revealed that the ERK1/2 target, mediator 1 (MED1), is recruited to the CYP24, but not the TRPV6, promoter following 1,25(OH)2D3 treatment. MED1 phosphorylation was sensitive to activators and inhibitors of the ERK1/2 signaling and MED1 siRNA reduced 1,25(OH)2D3-regulated human CYP24 promoter activity. This suggests ERK1/2 signaling enhances 1,25(OH)2D3 effects on the CYP24 promoter by MED1-mediated events. Our data show that there are both promoter-specific and cell stage-specific roles for the ERK signaling pathway on 1,25(OH)2D3-mediated gene induction in enterocyte-like Caco-2 cells.
登录
查看更多内容
影响因子：
4.8
作者：
Kerry, DM;Dwivedi, PP;May, BK
通讯作者：
May, BK
影响因子：
4.7
作者：
BELLELI, A;LEVY, J;LAMPRECHT, SA
通讯作者：
LAMPRECHT, SA
影响因子：
4.8
作者：
Bastien, J;Adam-Stitah, S;Rochette-Egly, C
通讯作者：
Rochette-Egly, C
DOI：
10.1016/j.jsbmb.2006.12.021
发表时间：
2007-03-01
影响因子：
4.1
作者：
Arriagada, Gloria;Paredes, Roberto;Montecino, Martin
通讯作者：
Montecino, Martin
影响因子：
--
作者：
Barletta, F;Freedman, LP;Christakos, S
通讯作者：
Christakos, S