The study of effects of amelogenin splice isoforms on the differentiation of mesenchymal cells

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。与对照组相比，在培养的第14天和第21天，釉原蛋白的加入分别增加了碱性磷酸酶活性和糖胺聚糖的分泌。定量PCR（Q-PCR）分析显示，LRAP在分化7天时增加Runx 2、Col 2a 1和聚集蛋白聚糖的基因表达水平。此外，M180和LRAP在培养28天时均显著增加ALP、Aggrecan、Col 10a 1和骨桥蛋白的基因表达水平。溴脱氧尿苷测定和Wnt信号的Q-PCR分析表明，M180和LRAP都降低了细胞增殖，但可能通过改变非经典Wnt信号诱导细胞分化。

We examined the effects of M180 and LRAP on a chondrogenic cell line, ATDC5, to investigate the role of amelogenins in chondrogenesis. The 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 of Runx2, Col2a1 and Aggrecan at 7 days of differentiation. Moreover, both M180 and LRAP significantly increased the gene expression levels of ALP, Aggrecan, Col10a1 and osteopontin at 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.