Amelogenin splice isoforms stimulate chondrogenic differentiation of ATDC5 cells.
Amelogenin splice isoforms stimulate chondrogenic differentiation of ATDC5 cells.
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Amelogenin 剪接亚型刺激 ATDC5 细胞的软骨分化。
DOI：
10.1111/j.1601-0825.2012.01967.x
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Igarashi K.
中科院分区：
文献类型：
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作者：
Mitani K;Haruyama N;Hatakeyama J;Igarashi K.
釉原蛋白是牙釉质中最丰富的基质蛋白。在釉原蛋白亚型中，全长釉原蛋白（M180）和富含亮氨酸的釉原蛋白肽（LRAP）在各种组织中表达，并被认为是间充质细胞中的信号分子。在这里，我们研究了M180和LRAP对软骨形成细胞系ATDC 5的影响，以研究釉原蛋白在软骨形成中的作用。结果釉原蛋白的加入增加了ATDC 5细胞的碱性磷酸酶活性和糖胺聚糖的分泌，与对照组相比，分别为14和21天。定量PCR（Q-PCR）分析显示，LRAP在分化7天时增加Runx 2、Col 2a 1和Aggrecan 3个基因的表达水平。此外，M180和LRAP在培养28天时均显著增加ALP、Aggrecan、Col 10a 1和骨桥蛋白基因的表达水平。结论M180和LRAP均能促进ATDC 5细胞向软骨细胞的分化和成熟。
ObjectiveAmelogenins are the most abundant matrix proteins in enamel. Among the amelogenin isoforms, full‐length amelogenin (M180) and leucine‐rich amelogenin peptide (LRAP) are expressed in various tissues and are implicated as signalling molecules in mesenchymal cells. Here, we examined the effects of M180 and LRAP on a chondrogenic cell line, ATDC5, to investigate the role of amelogenins in chondrogenesis.Materials and MethodsRecombinant mouse M180‐ or LRAP‐protein‐containing medium or control medium was mixed with a chondrogenesis‐stimulating medium, and changes in the phenotype, gene expression levels and cell proliferation of cultured ATDC5 cells were analysed.ResultsThe addition of amelogenins increased alkaline phosphatase activity and glycosaminoglycan secretion at 14 and 21 days of culture, respectively, as compared with the control. Quantitative PCR (Q‐PCR) analysis revealed that LRAP increased the gene expression levels ofRunx2,Col2a1andAggrecanat 7 days of differentiation. Moreover, both M180 and LRAP significantly increased the gene expression levels ofALP,Aggrecan,Col10a1andosteopontinat 28 days of culture. Bromodeoxyuridine assay and Q‐PCR analysis for Wnt signalling indicated that both M180 and LRAP reduced proliferation, but induced the cell differentiation possibly through altered non‐canonical Wnt signalling.ConclusionM180 and LRAP accelerate chondrogenic differentiation and maturation of ATDC5 cells.