Oligonucleotide array analysis of cyclic tension-responsive genes in human periodontal ligament fibroblasts.
Oligonucleotide array analysis of cyclic tension-responsive genes in human periodontal ligament fibroblasts.
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DOI：
10.1016/j.biocel.2007.01.015
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发表时间：
2007
期刊：
影响因子：
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通讯作者：
K. Yamashiro;F. Myokai;K. Hiratsuka;Tadashi Yamamoto;Kyoko Senoo;H. Arai;F. Nishimura;Y. Abiko
中科院分区：
文献类型：
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作者：
K. Yamashiro;F. Myokai;K. Hiratsuka;Tadashi Yamamoto;Kyoko Senoo;H. Arai;F. Nishimura;Y. Abiko
机械压力导致差异基因表达，这是将刺激转化为生化信号的关键。在咬合力等生理应激条件下，人牙周膜成纤维细胞(HPLF)与牙周组织的动态平衡有关，但对机械转导的反应变化尚不明确。我们假设循环张力反应(CT)基因可能被用来识别一系列机械转导的基本途径。我们的目标是对培养的HPLF中的CT基因进行分类。对HPLF进行长达16h的循环拉伸，并从张力负荷和静态HPLF中提取总RNA。寡核苷酸阵列分析显示，122个CT基因的mRNA积累发生了显著变化，并用K-均值聚类法对其动力学进行了分配。利用50个CT基因完成了HPLF机械转导的独创性路径分析。这一分析表明，循环张力立即下调所有核转录因子，除了v-FOS FBJ作为早期反应基因反应的小鼠骨肉瘤病毒癌基因同源基因(FOS)。反过来，转录因子如肿瘤蛋白P53结合蛋白2(TP53BP2)和核外分子如肾上腺素能受体β2(ADRB2)在1-2小时后上调，这可能导致基本的高密度脂蛋白功能以适应循环张力。随后使用Rho相关激酶(ROCK)的药物抑制剂Y27632进行的抑制分析表明，HPLF既有ROCK依赖的CT基因，也有ROCK非依赖的CT基因。机械应力可以影响许多基因的表达，特别是早期反应基因的表达；FOS启动了HPLF行为的改变，以控制牙周膜的动态平衡。
Mechanical stress results in differential gene expression that is critical to convert the stimulus into biochemical signals. Under physiological stress such as occlusal force, human periodontal ligament fibroblasts (HPLF) are associated with homeostasis of periodontal tissues however the changes in response to mechanotransduction remain uncharacterized. We hypothesized that cyclic tension-responsive (CT) genes may be used to identify a set of fundamental pathways of mechanotransduction. Our goal was to catalogue CT genes in cultured HPLF. HPLF were subjected to cyclic tension up to 16h, and total RNA was isolated from both tension-loaded and static HPLF. The oligonucleotide arrays analysis revealed significant changes of mRNA accumulation for 122 CT genes, and their kinetics were assigned by the K-means clustering methods. Ingenuity Pathway Analysis was completed for HPLF mechanotransduction using 50 CT genes. This analysis revealed that cyclic tension immediately down-regulated all nuclear transcription factors except v-fos FBJ murine osteosarcoma viral oncogene homolog (FOS) reacting as an early responsive gene. In turn, transcription factors such as tumor protein p53 binding protein 2 (TP53BP2), and extra-nuclear molecules such as adrenergic receptor β2 (ADRB2) were up-regulated after 1–2h, which may result in fundamental HPLF functions to adapt to cyclic tension. Subsequent inhibition assays using Y27632, a pharmacologic inhibitor of Rho-associated kinase (ROCK), suggested that HPLF has both ROCK-dependent and ROCK-independent CT genes. Mechanical stress was found to effect the expression of numerous genes, in particular, expression of an early responsive gene; FOS initiates alteration of HPLF behaviors to control homeostasis of the periodontal ligament.