Relationship of bone morphogenetic protein expression during osteoblast differentiation to wild type p53.
Relationship of bone morphogenetic protein expression during osteoblast differentiation to wild type p53.
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成骨细胞分化过程中骨形态发生蛋白表达与野生型p53的关系。
DOI：
10.1016/j.orthres.2005.04.010.1100230616
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Kolman,Kevin
中科院分区：
文献类型：
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作者：
Chandar,Nalini;Swindle,John;Szajkovics,Ann;Kolman,Kevin
我们以前已经表明p53在成骨细胞分化中具有特定的作用，通过其调节某些骨特异性蛋白表达的能力。在这项研究中，我们发现成骨细胞骨肉瘤细胞中的矿化基质形成与野生型p53的存在直接相关。为了进一步了解p53在分化中的重要性，我们研究了成骨细胞分化过程中p53与骨形态发生蛋白（BMP）（BMP 1，2，3A，3B（GDF-10），4，5，6，7，8A和8B）之间的关系。用RNase保护实验检测几种骨形成蛋白在分化型ROS 17/2.8成骨肉瘤细胞中的表达。BMPs 1、2、3a、3b和7的表达在体外分化过程中呈时间依赖性调节。为了确定p53是否在此过程中发挥作用，我们使用了稳定表达温度敏感性p53的鼠骨肉瘤细胞系。将细胞暴露于抗坏血酸和甘油磷酸盐以加速体外成骨细胞分化，并保持在32或37 °C下以表达野生型或突变型p53表型。BMP-2、BMP-4和BMP-7的表达以p53依赖性方式调节。我们能够通过RT-PCR独立地证实BMP-2的p53依赖性。虽然BMP-2表达在野生型和突变型p53的存在下都是明显的，但仅在表达野生型p53的细胞中观察到受调节的表达。野生型p53的瞬时过表达没有导致与稳定表达相同的BMP-2反应，表明p53的存在可能对成骨细胞分化的有序发展很重要，而不是对基因表达的直接影响。p53和这些骨特异性标志物之间的功能关系进行了讨论。© 2005骨科研究学会。由爱思唯尔有限公司出版。保留所有权利。
We have previously shown p53 to have a specific role in osteoblast differentiation by its ability to regulate expression of certain bone specific proteins. In this study, we show mineralized matrix formation in vivo to be directly related to the presence of wild type p53 in osteoblastic osteosarcoma cells. In order to further understand the importance of p53 in differentiation, we investigated the relationship between p53 and Bone Morphogenetic Proteins (BMPs) (BMP 1, 2, 3A, 3B (GDF‐10), 4, 5, 6, 7, 8A and 8B) during osteoblast differentiation. The expression of several BMPs were tested using RNase Protection Assay in differentiating ROS17/2.8 osteoblastic osteosarcoma cells. The expression of BMPs 1, 2, 3a, 3b and 7 showed time dependent modulation during in vitro differentiation. In order to determine if p53 has a role in this process, we used a murine osteosarcoma cell line stably expressing a temperature sensitive p53. Cells were exposed to ascorbic acid and glycerophosphates to hasten in vitro osteoblast differentiation and maintained either at 32 or 37 °C for expression of the wild type or mutant p53 phenotype. The expression of BMP‐2, BMP‐4 and BMP‐7 were modulated in a p53 dependent fashion. We were able to confirm the p53 dependency of BMP‐2 independently by RT‐PCR. While BMP‐2 expression was evident in the presence of both wild type and mutant p53, regulated expression was seen only in cells expressing wild type p53. Transient over expression of wild type p53 did not result in the same BMP‐2 response as stable expression showing that the presence of p53 may be important for an orderly development of osteoblast differentiation rather than a direct effect on gene expression. The functional relationship between p53 and these bone specific markers is discussed. © 2005 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.