Noggin Prevents Osteogenesis but Induces Chondrogenesis in a Human Mesenchymal Cell Line (USAC)

Y. Yoshizawa, K. Yagami, Yohei Uyama, S. Kakuta, M. Nagumo
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引用次数: 1

Abstract

The differentiation of mesenchymal stem cells (MSCs) into chondrocytes and osteoblasts is minutely regulated by bone morphogenetic proteins (BMPs) and various factors . There is no study describing the role of noggin in the differentiation from MSCs to chondrocytes or osteoblasts. We thus examined the effect of noggin onchondrogenesis and osteogenesis in a human cell line that expresses chondrocytic phenotypes, and the role of noggin in these processes . Noggin induces chondrogenic differentiation, and its repression induces osteogenic differentiation through BMP effects. Noggin is reported to bind several BMPs and to inhibit their functions, but its exact role is not yet clarified. USAC cells usually show chondrogenic phenotypes and also have the potential to differentiate into osteoblasts and adipocytes under somecircumstances . We investigated the effects of noggin on chondrogenic and osteogenic differentiationof USAC cells in vitro and in vivo. After USAC cells were treated with noggin antisense oligonucleotide (As-Noggin) or noggin protein (rhNoggin), RT-PCR was performed to detect mRNAsof type I collagen (Col I), type II collagen (Col II), Cbfal, aggrecan (AG), Sox9 and osteocalcin (OC) on days 3, 7 and 14. Western blot analysis was done to detect Col II, BMP-2, BMP-4, Cbfal and OC proteins . Toluidine blue staining and immunostaining for Col II, BMP-2 , Cbfal andOC were also performed. As-Noggin induced the up-regulation of Col I and OC mRNAexpression in USAC cells time-dependently, while it down-regulated the expressionof Col II and AG mRNAs . As-Noggin stimulated BMP-2, Cbfal and OC production, but it reduced the production of Col II and BMP-4 . Moreover, OC mRNA expression and BMP-2 production were down-regulated by the addition of rhNoggin. The same results were obtained by immunostaining. These results suggest that noggin may regulate chondrogenic and osteogenic differentiation through the production of BMP-2 and BMP-4.
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Noggin在人间充质细胞系(USAC)中阻止骨形成但诱导软骨形成
间充质干细胞(MSCs)向软骨细胞和成骨细胞的分化受骨形态发生蛋白(BMPs)和多种因素的精细调控。目前还没有研究描述noggin在MSCs向软骨细胞或成骨细胞分化中的作用。因此,我们研究了noggin在表达软骨细胞表型的人类细胞系中对软骨形成和成骨的影响,以及noggin在这些过程中的作用。Noggin诱导软骨分化,其抑制作用通过BMP诱导成骨分化。据报道,Noggin与几种bmp结合并抑制其功能,但其确切作用尚不清楚。USAC细胞通常表现为软骨细胞表型,在某些情况下也有分化成成骨细胞和脂肪细胞的潜力。我们在体外和体内研究了noggin对USAC细胞成软骨和成骨分化的影响。用头蛋白反意义寡核苷酸(As-Noggin)或头蛋白(rhNoggin)处理USAC细胞后,于第3、7、14天采用RT-PCR检测I型胶原(Col I)、II型胶原(Col II)、Cbfal、聚集蛋白(AG)、Sox9和骨钙素(OC)的mrna。Western blot检测Col II、BMP-2、BMP-4、Cbfal和OC蛋白。对Col II、BMP-2、Cbfal和doc进行甲苯胺蓝染色和免疫染色。As-Noggin诱导USAC细胞中Col I和OC mrna的表达呈时间依赖性上调,而下调Col II和AG mrna的表达。As-Noggin刺激了BMP-2、Cbfal和OC的生成,但降低了Col II和BMP-4的生成。此外,添加rhNoggin可下调OC mRNA的表达和BMP-2的产生。免疫染色也得到同样的结果。这些结果表明,noggin可能通过产生BMP-2和BMP-4来调节软骨和成骨分化。
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