Institute of Endocrinology and Metabolism, The Second Xiang-Ya Hospital of Central South University, Changsha, Hunan 410011, China ; Department of Endocrinology and Metabolism, Xiang-Ya Hospital of Central South University, Changsha, Hunan 410008, China.
Int J Endocrinol. 2013;2013:679763. doi: 10.1155/2013/679763. Epub 2013 Mar 19.
Wisp3 gene mutation was shown to cause spondyloepiphyseal dysplasia tarda with progressive arthropathy (SRDT-PA), but the underlying mechanism is not clear. To clarify this mechanism, we constructed the wild and mutated Wisp3 expression vectors and transfected into human chondrocytes lines C-20/A4; Wisp3 proteins subcellular localization, cell proliferation, cell apoptosis, and Wisp3-mediated gene expression were determined, and dynamic secretion of collagen in transfected chondrocytes was analyzed by (14)C-proline incorporation experiment. Mutated Wisp3 protein increased proliferation activity, decreased apoptosis of C-20/A4 cells, and aggregated abnormally in cytoplasm. Expression of collagen II was also downregulated in C-20/A4 cells transfected with mutated Wisp3. Wild type Wisp3 transfection increased intracellular collagen content and extracellular collagen secretion, but the mutated Wisp3 lost this function, and the peak phase of collagen secretion was delayed in mutated Wisp3 transfected cells. Thus abnormal protein distribution, cell proliferation, collagen synthesis, and secretion in Wisp3 mutated chondrocytes might contribute to the pathogenesis of SEDT-PA.
Wisp3 基因突变可导致迟发性脊椎骨骺发育不良伴进行性关节炎(SRDT-PA),但其发病机制尚不清楚。为阐明这一机制,我们构建了野生型和突变型 Wisp3 表达载体,并转染人软骨细胞系 C-20/A4;检测 Wisp3 蛋白的亚细胞定位、细胞增殖、细胞凋亡以及 Wisp3 介导的基因表达,并通过(14)C-脯氨酸掺入实验分析转染软骨细胞中胶原的动态分泌。突变型 Wisp3 蛋白增加了 C-20/A4 细胞的增殖活性,降低了细胞凋亡,并在细胞质中异常聚集。转染突变型 Wisp3 的 C-20/A4 细胞中 II 型胶原的表达也下调。野生型 Wisp3 转染增加了细胞内胶原含量和细胞外胶原分泌,但突变型 Wisp3 丧失了这种功能,并且突变型 Wisp3 转染细胞中胶原分泌的峰值阶段延迟。因此,Wisp3 突变型软骨细胞中异常的蛋白分布、细胞增殖、胶原合成和分泌可能导致 SEDT-PA 的发病机制。