Identification of mechanosensitive genes in osteoblasts by comparative microarray studies using the rotating wall vessel and the random positioning machine
Identification of mechanosensitive genes in osteoblasts by comparative microarray studies using the rotating wall vessel and the random positioning machine
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DOI：
10.1002/jcb.21218
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发表时间：
2007-06-01
影响因子：
4
通讯作者：
Jo, Hanjoong
中科院分区：
文献类型：
--
作者：
Patel, Mamta J.;Liu, Wenbin;Jo, Hanjoong
太空飞行的失重或微重力诱导骨丢失，部分原因是由于未知机制导致的骨形成减少。由于在太空中进行实验的困难，一些地面模拟器，如旋转壁容器（RWV）和随机定位机（RPM）已成为继续研究空间生物学的重要场所。然而，这些模拟器还没有系统地相互比较或机械刺激模型。在这里，我们假设暴露于RWV抑制分化和改变2 T3细胞的基因表达谱，这些机械敏感基因的一个子集的行为方式一致的RPM和相反的趋势所引起的机械刺激小鼠胸腺。2 T3前成骨细胞暴露于RWV 3天抑制碱性磷酸酶活性，分化的标志物，并下调61和上调45个基因的两倍以上相比，静态1克控制，如微阵列分析所示。微阵列结果通过实时PCR和/或蛋白质印迹法确认了7个独立的基因和蛋白质，包括骨调节蛋白、runx 2和骨甘氨酸。将RWV数据与我们先前发表的RPM微阵列研究进行比较，发现有14个机械敏感基因发生了相同方向的变化。将RWV和RPM结果与来自独立小组报告的机械加载小鼠tibodies的微阵列数据进行进一步比较，发现包括骨甘素在内的三个基因被加载上调，并被我们的模拟器下调。这些机械敏感基因可能提供新的见解，了解调节骨形成的机制和潜在的目标，对减少骨形成的对策，在太空飞行和病理学与缺乏骨形成。
Weightlessness or microgravity of spaceflight induces bone loss due in part to decreased bone formation by unknown mechanisms. Due to difficulty in performing experiments in space, several ground-based simulators such as the Rotating Wall Vessel (RWV) and Random Positioning Machine (RPM) have become critical venues to continue studying space biology. However, these simulators have not been systematically compared to each other or to mechanical stimulating models. Here, we hypothesized that exposure to RWV inhibits differentiation and alters gene expression profiles of 2T3 cells, and a subset of these mechanosensitive genes behaves in a manner consistent to the RPM and opposite to the trends incurred by mechanical stimulation of mouse tibiae. Exposure of 2T3 preosteoblast cells to the RWV for 3 days inhibited alkaline phosphatase activity, a marker of differentiation, and downregulated 61 and upregulated 45 genes by more than twofold compared to static 1 g controls, as shown by microarray analysis. The microarray results were confirmed by real-time PCR and/or Western blots for seven separate genes and proteins including osteomodulin, runx2, and osteoglycin. Comparison of the RWV data to the RPM microarray study that we previously published showed 14 mechanosensitive genes that changed in the same direction. Further comparison of the RWV and RPM results to microarray data from mechanically loaded mouse tibiae reported by an independent group revealed that three genes including osteoglycin were upregulated by the loading and downregulated by our simulators. These mechanosensitive genes may provide novel insights into understanding the mechanisms regulating bone formation and potential targets for countermeasures against decreased bone formation during space flight and in pathologies associated with lack of bone formation.