Effects of CTGF/Hcs24, a product of a hypertrophic chondrocyte-specific gene, on the proliferation and differentiation of chondrocytes in culture
Effects of CTGF/Hcs24, a product of a hypertrophic chondrocyte-specific gene, on the proliferation and differentiation of chondrocytes in culture
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DOI：
10.1210/endo.141.1.7267
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发表时间：
2000-01-01
期刊：
影响因子：
4.8
通讯作者：
Takigawa, M
中科院分区：
文献类型：
--
作者：
Nakanishi, T;Nishida, T;Takigawa, M
最近，我们从人软骨肉瘤来源的软骨细胞系HCS-2/8中克隆了一个主要在软骨细胞中表达的信使RNA（mRNA），并发现其基因，命名为hcs 24，与结缔组织生长因子（CTGF）的基因相同。本实验研究了CTGF/Hcs 24在软骨细胞系HCS-2/8和兔生长软骨（RGC）细胞中的功能。用产生CTGF/Hcs 24正义RNA（mRNA）的重组腺病毒转染的HCS-2/8细胞比用对照腺病毒转染的HCS-2/8细胞增殖更快。重组腺病毒介导CTGF/Hcs 24正义RNA转染的HCS-2/8细胞表达的聚集蛋白聚糖和X型胶原的mRNA水平高于对照细胞。为了阐明CTGF/Hcs 24对细胞的直接作用，我们用CTGF/Hcs 24表达载体转染HeLa细胞，获得稳定的转染子，并从转染子的条件培养基中纯化重组CTGF/Hcs 24蛋白。重组CTGF/Hcs 24以剂量依赖性方式有效地促进HCS-2/8细胞和RGC细胞的增殖，并以剂量依赖性方式增加这些细胞中蛋白多糖的合成。此外，CTGF/Hcs 24的这些刺激作用通过加入抗CTGF抗体而被中和。此外，重组CTGF/Hcs 24有效地增加培养的RGC细胞中的碱性磷酸酶活性。此外，RT-PCR分析显示，重组CTGF/Hcs 24刺激培养的RGC细胞中聚集蛋白聚糖以及II型和X型胶原的基因表达。这些结果表明，CTGF/Hcs 24直接促进软骨细胞的增殖和分化。
Recently, we cloned a messenger RNA (mRNA) predominantly expressed in chondrocytes from a human chondrosarcoma-derived chondrocytic cell line, HCS-2/8, by differential display PCR and found that its gene, named hcs24, was identical with that of connective tissue growth factor (CTGF). Here we investigated CTGF/Hcs24 func- tion in the chondrocytic cell line HCS-2/8 and rabbit growth cartilage (RGC) cells. HCS-2/8 cells transfected with recombinant adenoviruses that generate CTGF/Hcs24 sense RNA (mRNA) proliferated more rapidly than HCS-2/8 cells transfected with control adenoviruses. HCS-2/8 cells transfected with recombinant adenoviruses that generate CTGF/Hcs24 sense RNA expressed more mRNA of aggrecan and type X collagen than the control cells. To elucidate the direct action of CTGF/Hcs24 on the cells, we transfected HeLa cells with CTGF/Hcs24 expression vectors, obtained stable transfectants, and purified recombinant CTGF/Hcs24 protein from conditioned medium of the transfectants. The recombinant CTGF/Hcs24 effectively promoted the proliferation of HCS-2/8 cells and RGC cells in a dose-dependent manner and also dose dependently increased proteoglycan synthesis in these cells. In addition, these stimulatory effects of CTGF/Hcs24 were neutralized by the addition of anti-CTGF antibodies. Furthermore, the recombinant CTGF/Hcs24 effectively increased alkaline phosphatase activity in RGC cells in culture. Moreover, RT-PCR analysis revealed that the recombinant CTGF/Hcs24 stimulated gene expression of aggrecan and collagen types II and X in RGC cells in culture. These results indicate that CTGF/Hcs24 directly promotes the proliferation and differentiation of chondrocytes.