Zhang Xiaoxue, Dowd Diane R, Moore Meika C, Kranenburg Tanya A, Meester-Smoor Magda A, Zwarthoff Ellen C, MacDonald Paul N
Department of Pharmacology, Case Western Reserve University, Cleveland, Ohio 44106, USA.
J Biol Chem. 2009 Jul 3;284(27):18174-83. doi: 10.1074/jbc.M109.001354. Epub 2009 Apr 22.
The vitamin D endocrine system is essential for calcium and phosphate homeostasis and skeletal mineralization. The 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) hormone binds to the vitamin D receptor (VDR) to regulate gene expression. These gene products in turn mediate the actions of 1,25(OH)(2)D(3) in mineral-regulating target cells such as the osteoblast. We showed previously that meningioma 1 (MN1) is a novel target of 1,25(OH)(2)D(3) in MG-63 osteoblastic cells and that it is a coactivator for VDR-mediated transcription (Sutton, A. L., Zhang, X., Ellison, T. I., and MacDonald, P. N. (2005) Mol. Endocrinol. 19, 2234-2244). However, the functional significance of MN1 in osteoblastic cell biology is largely unknown. Here, we demonstrate that MN1 expression is increased dramatically during differentiation of primary osteoblastic cells. Using calvarial osteoblasts derived from wild-type and MN1 knock-out mice, we provide data supporting an essential role of MN1 in maintaining appropriate osteoblast proliferation, differentiation, and function. MN1 knock-out osteoblasts displayed altered morphology, decreased growth rate, impaired motility, and attenuated 1,25(OH)(2)D(3)/VDR-mediated transcription as well as reduced alkaline phosphatase activity and mineralized nodule formation. MN1 null osteoblasts were also impaired in supporting osteoclastogenesis in co-culture studies presumably because of marked reduction in the RANKL:OPG ratio in the MN1 null cells. Mechanistic studies supported a transcriptional role for MN1 in controlling RANKL gene expression through activation of the RANKL promoter. Cumulatively, these studies indicate an important role for MN1 in maintaining the appropriate maturation and function of calvarial osteoblasts.
维生素D内分泌系统对于钙和磷的稳态以及骨骼矿化至关重要。1,25-二羟基维生素D(3)(1,25(OH)₂D₃)激素与维生素D受体(VDR)结合以调节基因表达。这些基因产物进而介导1,25(OH)₂D₃在矿物质调节靶细胞(如成骨细胞)中的作用。我们之前表明,脑膜瘤1(MN1)是MG-63成骨细胞中1,25(OH)₂D₃的新靶点,并且它是VDR介导转录的共激活因子(萨顿,A.L.,张,X.,埃里森,T.I.,和麦克唐纳,P.N.(2005年)《分子内分泌学》19,2234 - 2244)。然而,MN1在成骨细胞生物学中的功能意义在很大程度上尚不清楚。在此,我们证明在原代成骨细胞分化过程中MN1表达显著增加。使用源自野生型和MN1基因敲除小鼠的颅骨成骨细胞,我们提供的数据支持MN1在维持适当的成骨细胞增殖、分化和功能中起重要作用。MN1基因敲除的成骨细胞表现出形态改变、生长速率降低、运动能力受损、1,25(OH)₂D₃/VDR介导的转录减弱以及碱性磷酸酶活性和矿化结节形成减少。在共培养研究中,MN1缺失的成骨细胞在支持破骨细胞生成方面也受损,这可能是因为MN1缺失细胞中RANKL:OPG比值显著降低。机制研究支持MN1通过激活RANKL启动子在控制RANKL基因表达中起转录作用。总的来说,这些研究表明MN1在维持颅骨成骨细胞的适当成熟和功能中起重要作用。