Epigenetic regulation of BMP2 by 1,25-dihydroxyvitamin D3 through DNA methylation and histone modification.
Epigenetic regulation of BMP2 by 1,25-dihydroxyvitamin D3 through DNA methylation and histone modification.
复制标题
DOI：
10.1371/journal.pone.0061423
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Favus MJ
中科院分区：
文献类型：
--
作者：
Fu B;Wang H;Wang J;Barouhas I;Liu W;Shuboy A;Bushinsky DA;Zhou D;Favus MJ
遗传性高钙尿石形成（GHS）大鼠具有增加的肠钙吸收、减少的肾小管钙重吸收和低骨量，所有这些至少部分地由升高的维生素D受体（VDR）组织水平介导。1，25-二羟维生素D3（1，25（OH）2D 3）和骨形态发生蛋白2（BMP 2）分别对骨代谢和骨形成的正常维持至关重要。骨细胞调节的复杂性表明GHS大鼠中这两种重要调节剂的潜在相互作用。在本研究中，VDR-1，25（OH）2D 3复合物抑制来自GHS和SD大鼠的骨髓基质细胞（BMSCs）和UMR-106细胞系中的BMP 2表达。我们使用染色质免疫沉淀（ChIP）试验，以确定VDR结合到几个潜在的结合位点内的BMP 2启动子区域之一。该负区域也介导抑制报告基因活性。采用DNA甲基转移酶抑制剂5-氮杂-2 ′-脱氧胞苷（DAC）和组蛋白去乙酰化酶抑制剂阿司他丁A（TSA），在体外研究了1，25（OH）2D 3下调BMSCs和UMR-106细胞BMP 2的分子机制。DAC和TSA都与1，25（OH）2D 3结合激活BMP 2表达。亚硫酸氢盐DNA焦磷酸测序显示，1，25（OH）2D 3完全高甲基化同一BMP 2启动子区域中的单个CpG位点，该区域由ChIP和报告基因测定鉴定。ChIP检测还显示，1，25（OH）2D 3可以增加组蛋白抑制性标记H3 K9 me 2，并减少组蛋白H3在同一BMP 2启动子区域的乙酰化。综上所述，我们的结果表明，1，25（OH）2D 3与VDR结合，通过与BMP 2启动子区结合，下调BMSC和成骨细胞样UMR-106细胞中BMP 2基因的表达。这种1，25（OH）2D 3诱导的BMP 2转录抑制的机制涉及DNA甲基化和组蛋白修饰。该研究提供了新的证据，表明1，25（OH）2D 3通过下调体内和体外BMP 2表达来抑制骨形成。
Genetic hypercalciuric stone-forming (GHS) rats have increased intestinal Ca absorption, decreased renal tubule Ca reabsorption and low bone mass, all of which are mediated at least in part by elevated tissue levels of the vitamin D receptor (VDR). Both 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and bone morphogenetic protein 2 (BMP2) are critical for normal maintenance of bone metabolism and bone formation, respectively. The complex nature of bone cell regulation suggests a potential interaction of these two important regulators in GHS rats. In the present study, BMP2 expression is suppressed by the VDR-1,25(OH)2D3 complex in Bone Marrow Stromal Cells (BMSCs) from GHS and SD rat and in UMR-106 cell line. We used chromatin immunoprecipitation (ChIP) assays to identify VDR binding to only one of several potential binding sites within the BMP2 promoter regions. This negative region also mediates suppressor reporter gene activity. The molecular mechanisms underlying the down-regulation of BMP2 by 1,25(OH)2D3 were studied in vitro in BMSCs and UMR-106 cells using the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine (DAC) and the histone deacetylase inhibitor trichostatin A (TSA). Both DAC and TSA activate BMP2 expression in combination with 1,25(OH)2D3. Bisulfite DNA pyrosequencing reveals 1,25(OH)2D3 to completely hypermethylate a single CpG site in the same BMP2 promoter region identified by the ChIP and reporter gene assays. ChIP assays also show that 1,25(OH)2D3 can increase the repressive histone mark H3K9me2 and reduce the acetylation of histone H3 at the same BMP2 promoter region. Taken together, our results indicate that 1,25(OH)2D3 binding to VDR down-regulates BMP2 gene expression in BMSCs and osteoblast-like UMR-106 cells by binding to the BMP2 promoter region. The mechanism of this 1,25(OH)2D3-induced transcriptional repression of BMP2 involves DNA methylation and histone modification. The study provides novel evidence that 1,25(OH)2D3 represses bone formation through down-regulating BMP2 expression both in vivo and in vitro.
登录
查看更多内容
影响因子：
8.2
作者：
Jurutka, P W;Whitfield, G K;Haussler, M R
通讯作者：
Haussler, M R
影响因子：
4.8
作者：
Ecarot, B;Desbarats, M
通讯作者：
Desbarats, M
影响因子：
4.8
作者：
Hines, ER;Kolek, OI;Ghishan, FK
通讯作者：
Ghishan, FK
影响因子：
3.7
作者：
Drissi, H;Pouliot, A;van Wijnen, AJ
通讯作者：
van Wijnen, AJ
影响因子：
6.2
作者：
Karnauskas, AJ;van Leeuwen, JPTM;Favus, MJ
通讯作者：
Favus, MJ