Loss of MMP-2 in murine osteoblasts upregulates osteopontin and bone sialoprotein expression in a circuit regulating bone homeostasis.
Loss of MMP-2 in murine osteoblasts upregulates osteopontin and bone sialoprotein expression in a circuit regulating bone homeostasis.
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DOI：
10.1242/dmm.007914
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发表时间：
2013-03
影响因子：
4.3
通讯作者：
Martignetti JA
中科院分区：
文献类型：
--
作者：
Mosig RA;Martignetti JA
多中心骨溶解伴关节病（MOA; MIM 605156）是一种遗传性骨溶解和关节炎综合征，由基质金属蛋白酶2（MMP-2）丢失引起。我们最近证明，Mmp 2-/-小鼠代表了研究人类疾病的独特模型，具有人类综合症的许多特征，包括骨骼发育不良和成骨细胞行为缺陷。因此，我们试图探索MMP-2损失的次级分子效应，其与潜在的骨骼和成骨细胞表型共存。我们使用定量实时RT-PCR（qRT-PCR）通过Mmp 2 −/−和Mmp 2 +/+小鼠骨髓基质细胞（BMSC）的体外成骨细胞分化来测量成骨细胞相关基因的表达。采用免疫印迹法检测血清骨桥蛋白（OPN）水平，免疫组化法检测骨表达。使用siRNA抑制SaOS 2细胞中MMP-2的表达，并且分别通过qRT-PCR和western blot证实MMP-2在RNA和蛋白质水平上的表达降低。与对照组相比，Mmp 2 −/− BMSC诱导分化为成骨细胞显示出显著上调OPN和骨唾液蛋白（BSP）表达水平。Mmp 2 −/−小鼠血清和骨中OPN水平升高证明了体内转录上调得以维持。这些作用是可推广的，因为siRNA介导的抑制在培养的细胞中也上调OPN和BSP。已知OPN和BSP影响MMP-2的表达和活性，但先前未显示受MMP-2调节。这种新定义的电路的鉴定提供了对MOA表型潜在分子景观的深入了解，并突出了可能在正常骨稳态中发挥作用的途径。
Multicentric osteolysis with arthropathy (MOA; MIM 605156) is an inherited osteolyses and arthritis syndrome resulting from loss of matrix metalloproteinase 2 (MMP-2). We recently demonstrated that Mmp2–/– mice represent a unique model for the study of the human disease, sharing many features of the human syndrome including skeletal dysplasia and defects in osteoblast behavior. We therefore sought to explore the secondary molecular effects of MMP-2 loss, which coexist with the underlying skeletal and osteoblast phenotypes. We used quantitative real-time RT-PCR (qRT-PCR) to measure osteoblast-related gene expression through ex vivo osteoblast differentiation of bone marrow stromal cells (BMSC) from Mmp2−/− and Mmp2+/+ mice. We used western blot to measure osteopontin (OPN) serum levels and immunohistochemical staining to examine bone expression. MMP-2 expression was inhibited in SaOS2 cells using siRNA, and decreased MMP-2 expression at both RNA and protein levels was confirmed by qRT-PCR and western blot, respectively. Mmp2−/− BMSC induced to differentiate into osteoblasts were shown to significantly upregulate OPN and bone sialoprotein (BSP) expression levels compared with controls. Transcriptional upregulation was maintained in vivo, as demonstrated by increased levels of OPN in serum and bone in Mmp2−/− mice. These effects are generalizable because siRNA-mediated inhibition in cultured cells also upregulated OPN and BSP. OPN and BSP are known to affect MMP-2 expression and activity but have not previously been shown to be regulated by MMP-2. Identification of this newly defined circuitry provides insight into the potential molecular landscape underlying the MOA phenotype and highlights a pathway that might play a role in normal bone homeostasis.
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影响因子：
4.1
作者：
Gordon, Jonathan A. R.;Tye, Coralee E.;Goldberg, Harvey A.
通讯作者：
Goldberg, Harvey A.
影响因子：
7
作者：
Dean, Richard A.;Overall, Christopher M.
通讯作者：
Overall, Christopher M.
影响因子：
3.5
作者：
Mosig, Rebecca A.;Dowling, Oonagh;Martignetti, John A.
通讯作者：
Martignetti, John A.
影响因子：
4.2
作者：
Shapses, SA;Cifuentes, M;Denhardt, DT
通讯作者：
Denhardt, DT
影响因子：
4.1
作者：
Sommer, B;Bickel, M;Wetterwald, A
通讯作者：
Wetterwald, A