A-674563 increases chondrocyte marker expression in cultured chondrocytes by inhibiting Sox9 degradation
A-674563 increases chondrocyte marker expression in cultured chondrocytes by inhibiting Sox9 degradation
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A-674563 通过抑制 Sox9 降解来增加培养软骨细胞中软骨细胞标记物的表达
DOI：
10.1016/j.bbrc.2017.11.180
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发表时间：
2018
影响因子：
3.1
通讯作者：
Tsumaki N.
中科院分区：
文献类型：
--
作者：
Kobayashi T;Fujita K;Kamatani T;Matsuda S;Tsumaki N.
自体软骨细胞植入术是治疗关节软骨损伤的一种有效方法。然而，由于单层培养中软骨细胞的扩张，导致软骨细胞特性的丧失，其益处有限。因此，需要增强软骨细胞特性的培养条件。我们筛选了5822种化合物，发现A-674563增强了小鼠原代软骨细胞中几种软骨细胞标记基因（包括col2a1、AcanandCol11a2）的转录。利用环己亚胺、MG132和巴菲霉素A1进行的实验表明，Sox9是通过泛素-蛋白酶体途径降解的，而A-674563抑制了这种降解，导致Sox9蛋白的量增加。RNA测序转录组分析显示，A-674563增加了编码泛素特异性肽酶29的基因的表达，该基因可以诱导蛋白质的去泛素化。虽然确切的机制仍有待确定，我们的研究结果表明，A-674563可以促进培养条件，扩大软骨细胞而不失去软骨细胞特性。
The implantation of autologous chondrocytes is a therapeutic treatment for articular cartilage damage. However, the benefits are limited due to the expansion of chondrocytes in monolayer culture, which causes loss of chondrocytic characters. Therefore, culture conditions that enhance chondrocytic characters are needed. We screened 5822 compounds and found that A-674563 enhanced the transcription of several chondrocyte marker genes, includingCol2a1,AcanandCol11a2, in mouse primary chondrocytes. Experiments using cycloheximide, MG132 and bafilomycin A1 have revealed that Sox9 is degraded through the ubiquitin-proteasome pathway and that A-674563 inhibits this degradation, resulting in larger amount of Sox9 protein. RNA sequencing transcriptome analysis showed that A-674563 increases the expression of the gene that encodes ubiquitin-specific peptidase 29, which is known to induce the deubiquitination of proteins. Although the precise mechanism remains to be determined, our findings indicated that A-674563 could contribute to culture conditions that expand chondrocytes without losing chondrocytic characters.