Surface adhesion-mediated regulation of chondrocyte-specific gene expression in the nontransformed RCJ 3.1C5.18 rat chondrocyte cell line.
Surface adhesion-mediated regulation of chondrocyte-specific gene expression in the nontransformed RCJ 3.1C5.18 rat chondrocyte cell line.
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非转化 RCJ 3.1C5.18 大鼠软骨细胞系中软骨细胞特异性基因表达的表面粘附介导调节。
DOI：
10.1002/jbmr.5650110812
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Hahn,TJ
中科院分区：
文献类型：
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作者：
McDougall,S;Fu,YH;Lowe,GN;Williams,A;Polendo,R;Benya,PD;Iida-Klein,A;Fang,MA;Hahn,TJ
最近的证据表明，骨关节炎和其他关节疾病中软骨细胞功能的降低可能是由于软骨细胞外基质（ECM）的调节信号改变所产生的软骨细胞去分化。然而，目前还没有适合于该过程研究的哺乳动物软骨细胞系系统。因此，我们研究了ECM生长条件对非转化大鼠RCJ 3.1C5.18（RCJ）软骨细胞系中分化软骨细胞表型表达标志物的影响，包括II型胶原表达、聚集蛋白聚糖产生、连接蛋白基因表达和甲状旁腺激素（PTH）受体数量。在塑料上单层生长的RCJ细胞表现出去分化表型，其特征在于扁平的细胞形态，通过胶原蛋白溴化氰肽的二维凝胶图谱电泳测定，I型胶原蛋白产量>80%，II型胶原蛋白产量<5%。此外，聚集蛋白聚糖的产量低，连接蛋白mRNA的表达没有达到可检测的水平。在复合I型胶原/琼脂糖（0.15%-0.8%）凝胶（CAG）表面上在最小附着条件下转移生长7天后，RCJ细胞形成圆形软骨细胞形态和分化的软骨细胞基因表达模式，产生79%的II型和8%的I型胶原。稳态I型和II型前胶原mRNA水平与胶原蛋白表达平行改变。在CAG上生长的细胞中，聚集蛋白聚糖产量增加6倍，聚集蛋白聚糖核心蛋白和连接蛋白mRNA水平均显著增加。此外，最大PTH刺激cAMP生成增加15倍，与PTH受体数量增加相关。因此，RCJ软骨细胞系对软骨细胞特异性基因表达的ECM调节高度敏感。
Recent evidence suggests that decreased chondrocyte function in osteoarthritis and other articular disorders may be due to chondrocyte dedifferentiation produced by altered regulatory signals from the cartilage extracellular matrix (ECM). However, there are currently no mammalian chondrocytic cell line systems adapted to the study of this process. We therefore examined the effects of ECM growth conditions on markers of differentiated chondrocytic phenotype expression in the nontransformed rat RCJ 3.1C5.18 (RCJ) chondrocyte cell line, including type II collagen expression, aggrecan production, link protein gene expression, and parathyroid hormone (PTH) receptor number. RCJ cells grown in monolayer on plastic exhibited a dedifferentiated phenotype characterized by flattened cell morphology, with >80% type I collagen and <5% type II collagen production, as determined by two‐dimensional gel mapping electrophoresis of collagen cyanogen bromide peptides. In addition, aggrecan production was low, and link protein mRNA was not expressed at detectable levels. After transfer to growth under minimal attachment conditions on the surface of a composite type I collagen/agarose (0.15%–0.8%) gel (CAG) for 7 days, RCJ cells developed a rounded, chondrocytic morphology and a pattern of differentiated, chondrocytic gene expression, with 79% type II and 8% type I collagen production. Steady‐state type I and type II procollagen mRNA levels were altered in parallel with collagen protein expression. In cells grown on CAG, aggrecan production increased 6‐fold, and there was a marked increase in both aggrecan core protein and link protein mRNA levels. In addition, maximal PTH‐stimulated cAMP generation increased 15‐fold in association with an increased PTH receptor number. Therefore, the RCJ chondrocyte cell line is highly sensitive to ECM regulation of chondrocyte‐specific gene expression.