INTERLEUKIN-1-BETA-MODULATED GENE-EXPRESSION IN IMMORTALIZED HUMAN CHONDROCYTES
INTERLEUKIN-1-BETA-MODULATED GENE-EXPRESSION IN IMMORTALIZED HUMAN CHONDROCYTES
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DOI：
10.1172/jci117595
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发表时间：
1994-12-01
影响因子：
15.9
通讯作者：
APPERLEY, JF
中科院分区：
文献类型：
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作者：
GOLDRING, MB;BIRKHEAD, JR;APPERLEY, JF
用编码猴病毒40大T抗原的载体转染幼年肋软骨细胞原代培养物，琼脂糖悬浮培养选择，建立了人软骨细胞的永生化。生成的稳定细胞系显示出软骨细胞形态，在含血清培养基中的单层培养中具有连续增殖能力（bbb80传代），并且在含胰岛素的血清替代品中表达编码软骨细胞特异性胶原II、IX和XI和蛋白聚糖的mrna。它们不表达X型胶原或versicmrna。这些细胞合成并分泌细胞外基质分子，与II型胶原蛋白、大蛋白聚糖（PG-H，聚集蛋白）和硫酸软骨素-4和硫酸软骨素-6的单克隆抗体反应。白细胞介素-1 β (IL-1 β)降低LI型胶原mRNA水平，增加胶原酶、基质溶解素和直接早期基因（egr-1 c-fos、c-jun和jun-B） mRNA水平。在瞬时转染实验中，这些细胞系还表达了含有II型胶原基因（COL2A1）调控序列（-577/+ 3428 bp）的报告基因构建物，IL-1 β抑制了50-80%的表达。这些结果表明，IL-1 β对软骨特异性调节的永生化人软骨细胞可以作为研究正常和病理修复机制的模型。
Immortalized human chondrocytes were established by transfection of primary cultures of juvenile costal chondrocytes with vectors encoding simian virus 40 large T antigen and selection in suspension culture over agarose. Stable cell lines were generated that exhibited chondrocyte morphology, continuous proliferative capacity (> 80 passages) in monolayer culture in serum-containing medium, and expression of mRNAs encoding chondrocyte-specific collagens II, IX, and XI and proteoglycans in an insulin-containing serum substitute. They did not express type X collagen or versican mRNA. These cells synthesized and secreted extracellular matrix molecules that were reactive with monoclonal antibodies against type II collagen, large proteoglycan (PG-H, aggrecan), and chondroitin-4- and chondroitin-6-sulfate. Interleukin-1 beta (IL-1 beta) decreased the levels of type LI collagen mRNA and increased the levels of mRNAs for collagenase, stromelysin, and immediate early genes (egr-1 c-fos, c-jun, and jun-B). These cell lines also expressed reporter gene constructs containing regulatory sequences (-577/+3,428 bp) of the type II collagen gene (COL2A1) in transient transfection experiments, and IL-1 beta suppressed this expression by 50-80%. These results show that immortalized human chondrocytes displaying cartilage-specific modulation by IL-1 beta can be used as a model for studying normal and pathological repair mechanisms.