| 鲍远,黄俊明,吴颖星,等.Yes相关蛋白通过Wnt/β-catenin信号通路调控软骨细胞Sox9表达的研究.骨科,2016,7(3): 195-200. |
| Yes相关蛋白通过Wnt/β-catenin信号通路调控软骨细胞Sox9表达的研究 |
| Yes-associated protein regulates the expression of Sox9 in chondrocytes by Wnt/β-catenin signaling pathway |
| 投稿时间：2015-12-11 |
| DOI：DOI：10.3969/j.issn.1674-8573.2016.03.012 |
| 中文关键词: 软骨细胞 细胞分化 Yes相关蛋白 信号传导 基因表达调控 |
| 英文关键词: Chondrocytes Cell differentiation Yes-associated protein Signal transduction Gene expression regulation |
| 基金项目:国家自然科学基金（81171696，81472082） |
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| 中文摘要: |
| 目的 研究Yes相关蛋白（Yes-associated protein, YAP）通过Wnt/β-catenin信号通路调控软骨细胞Sox9表达的机制。方法 用siRNA分别沉默YAP基因和Lats1基因来调控YAP的活性，通过Real-time PCR检测软骨特异性基因和Wnt/β-catenin信号通路下游基因的表达；通过Western Blot检测GSK3β的磷酸化水平、活化β-catenin的蛋白水平以及Sox9的蛋白水平；并通过阿利新蓝染色检测软骨细胞外基质的分泌情况。用siRNA沉默YAP基因后，再用氯化锂干预细胞，通过Western Blot检测Sox9的表达和GSK3β的磷酸化水平。结果 沉默YAP基因后，磷酸化GSK3β和活化β-catenin的蛋白水平减少，c-Myc和Nanog基因表达减少，Sox9、Col2和Aggrecan基因表达升高，并且软骨细胞分泌基质增多；沉默Lats1基因后，磷酸化GSK3β和活化β-catenin的蛋白水平增加，c-Myc和Nanog基因表达上调，Sox9、Col2和Aggrecan基因表达减少，并且软骨细胞分泌基质减少。此外，氯化锂可以阻断沉默YAP引起的Sox9表达上调。结论 YAP通过Wnt/β-catenin信号通路调控软骨细胞Sox9的表达。 |
| 英文摘要: |
| Objective To investigate the mechanisms of Sox9 regulated by Yes-associated protein (Yap) in chondrocytes via Wnt signaling pathway. Methods Yap or Lats1 was knocked down with small interfering RNA to regulate the activity of Yap. Genes related to Wnt/β-catenin signaling pathway and cartilage-specific genes were detected by Real-time PCR. Wnt/β-catenin signaling was determined by detection of p-GSK3β and active β-catenin via Western blotting and Sox9 was also detected by Western blotting. The cartilage-specific extracellular matrix was assessed by alcian blue staining. Chondrocytes with Yap knockdown were treated with lithium chloride, then p-GSK3β and Sox9 were detected by Western blotting. Results After knockdown of Yap, Wnt signaling pathway was inhibited, leading to increased expression of Sox9, Col2 and Aggrecan and enhanced secretion of the cartilage-specific extracellular matrix. After Lats1 was knocked down, the obtained result was opposite completely. Moreover, lithium chloride could block the up-regulation of Sox9 expression induced by knockdown of Yap. Conclusion Yap regulates the expression of Sox9 in chondrocytes by Wnt signaling pathway. |
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