Regulation of collagenase-3 and osteocalcin gene expression by collagen and osteopontin in differentiating MC3T3-E1 cells
Regulation of collagenase-3 and osteocalcin gene expression by collagen and osteopontin in differentiating MC3T3-E1 cells
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DOI：
10.1074/jbc.m202604200
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发表时间：
2002-07-05
影响因子：
4.8
通讯作者：
Partridge, NC
中科院分区：
文献类型：
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作者：
D'Alonzo, RC;Kowalski, AJ;Partridge, NC
胶原酶-3和骨钙素mRNA在成骨细胞分化的后期阶段表达最大。在这里，我们证明，胶原酶-3 mRNA的表达在分化MC 3 T3-E1细胞依赖于抗坏血酸的存在下，抑制在胶原合成抑制剂，3，4-脱氢脯氨酸的存在下，并刺激生长在胶原蛋白在抗坏血酸的情况下。瞬时转染研究表明，胶原酶-3启动子的活性在细胞分化过程中增加，需要抗坏血酸的存在。此外，我们发现，在分化MC 3 T3-E1细胞，胶原酶-3基因表达增加的存在下，抗骨桥蛋白单克隆抗体，结合附近的RGD基序的这种蛋白质，而骨钙素的表达被抑制。此外，设计用于阻断配体与α（v）整联蛋白亚基的相互作用的RGD肽模拟物化合物增加骨钙蛋白表达并抑制胶原酶-3表达，表明RGD肽模拟物在成骨细胞中启动某些α（v）整联蛋白信号传导。总之，这些研究表明，在成骨细胞分化过程中刺激胶原酶-3表达需要合成胶原基质，并且骨桥蛋白和α（v）整合素在成骨细胞分化过程中对胶原酶-3和骨钙素表达发挥不同的调节作用。
Both collagenase-3 and osteocalcin mRNAs are expressed maximally during the later stages of osteoblast differentiation. Here, we demonstrate that collagenase-3 mRNA expression in differentiating MC3T3-E1 cells is dependent upon the presence of ascorbic acid, is inhibited in the presence of the collagen synthesis inhibitor, 3,4-dehydroproline, and is stimulated by growth on collagen in the absence of ascorbic acid. Transient transfection studies show that collagenase-3 promoter activity increases during cell differentiation and requires the presence of ascorbic acid. Additionally, we show that, in differentiating MC3T3-E1 cells, collagenase-3 gene expression increases in the presence of an anti-osteopontin monoclonal antibody that binds near the RGD motif of this protein, whereas osteocalcin expression is inhibited. Furthermore, an RGD peptidomimetic compound, designed to block interaction of ligands to the alpha(v) integrin subunit, increases osteocalcin expression and inhibits collagenase-3 expression, suggesting that the RGD peptidomimetic initiates certain alpha(v) integrin signaling in osteoblastic cells. Overall, these studies demonstrate that stimulation of collagenase-3 expression during osteoblast differentiation requires synthesis of a collagenous matrix and that osteopontin and alpha(v) integrins exert divergent regulation of collagenase-3 and osteocalcin expression during osteoblast differentiation.