Regulation of chondrocyte gene expression by osteogenic protein-1
Regulation of chondrocyte gene expression by osteogenic protein-1
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DOI：
10.1186/ar3300
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发表时间：
2011-01-01
影响因子：
4.9
通讯作者：
Loeser, Richard F.
中科院分区：
文献类型：
--
作者：
Chubinskaya, Susan;Otten, Lori;Loeser, Richard F.
简介：本研究的目的是利用Affiliate基因芯片研究人关节软骨细胞中哪些基因受成骨蛋白-1（OP-1）的调控，以了解OP-1在软骨稳态中的作用。从12个正常踝关节软骨样本中酶促分离的软骨细胞在高密度单层中培养，并转染OP-1反义寡核苷酸在脂转染素存在下处理或用重组OP-1（100 ng/ml）处理48小时，然后分离RNA。基因表达谱分析采用HG-U133 A基因芯片。临界值选择为与对照的1.5倍差异。所选基因阵列结果通过实时PCR和蛋白聚糖合成和信号转导的体外测量进行验证。OP-1在多个水平上控制软骨稳态，包括调节负责软骨细胞骨架的基因（细胞周期蛋白D，Talin 1和细胞周期蛋白M1），基质产生和其他合成代谢途径（转化生长因子-β（TGF-β）/骨形态发生蛋白（BMP）、胰岛素样生长因子（IGF）、血管内皮生长因子（VEGF）、负责骨形成的基因等）以及细胞因子、神经介质、以及导致基质降解和细胞死亡的各种分解代谢途径。在许多这些情况下，OP-1调制的表达，不仅配体，而且其受体，介体的下游信号，激酶负责激活的途径，结合蛋白负责抑制的途径，和转录因子，诱导转录responses.Conclusions：基因阵列数据强烈表明OP-1在人类软骨稳态的关键作用。OP-1调节许多代谢途径，不仅限于其充分记录的合成代谢功能，还包括其抗分解代谢活性。了解OP-1在软骨中的功能将为OP-1蛋白作为软骨再生和修复的治疗性治疗的应用提供强有力的理由。
Introduction: The objective of this study was to investigate which genes are regulated by osteogenic protein-1 (OP-1) in human articular chondrocytes using Affimetrix gene array, in order to understand the role of OP-1 in cartilage homeostasis.Methods: Chondrocytes enzymatically isolated from 12 normal ankle cartilage samples were cultured in high-density monolayers and either transfected with OP-1 antisense oligonucleotide in the presence of lipofectin or treated with recombinant OP-1 (100 ng/ml) for 48 hours followed by RNA isolation. Gene expression profiles were analyzed by HG-U133A gene chips from Affimetrix. A cut-off was chosen at 1.5-fold difference from controls. Selected gene array results were verified by real-time PCR and by in vitro measures of proteoglycan synthesis and signal transduction.Results: OP-1 controls cartilage homeostasis on multiple levels including regulation of genes responsible for chondrocyte cytoskeleton (cyclin D, Talin1, and Cyclin M1), matrix production, and other anabolic pathways (transforming growth factor-beta (TGF-beta)/bone morphogenetic protein (BMP), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), genes responsible for bone formation, and so on) as well as regulation of cytokines, neuromediators, and various catabolic pathways responsible for matrix degradation and cell death. In many of these cases, OP-1 modulated the expression of not only the ligands, but also their receptors, mediators of downstream signaling, kinases responsible for an activation of the pathways, binding proteins responsible for the inhibition of the pathways, and transcription factors that induce transcriptional responses.Conclusions: Gene array data strongly suggest a critical role of OP-1 in human cartilage homeostasis. OP-1 regulates numerous metabolic pathways that are not only limited to its well-documented anabolic function, but also to its anti-catabolic activity. An understanding of OP-1 function in cartilage will provide strong justification for the application of OP-1 protein as a therapeutic treatment for cartilage regeneration and repair.