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首页> 外文期刊>Journal of cellular biochemistry. >Prolonged culture of HOS 58 human osteosarcoma cells with 1,25-(OH)(2)-D(3), TGF-beta, and dexamethasone reveals physiological regulation of alkaline phosphatase, dissociated osteocalcin gene expression, and protein synthesis and lack of mineralizati
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Prolonged culture of HOS 58 human osteosarcoma cells with 1,25-(OH)(2)-D(3), TGF-beta, and dexamethasone reveals physiological regulation of alkaline phosphatase, dissociated osteocalcin gene expression, and protein synthesis and lack of mineralizati

机译：用1,25-（OH）（2）-D（3），TGF-β和地塞米松长时间培养HOS 58人骨肉瘤细胞显示出生理调节碱性磷酸酶，解离骨钙素基因表达，蛋白质合成和缺乏矿物质

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Cultured rodent osteoblastic cells reiterate the phenotypic differentiation and maturation of osteoblasts seen in vivo. As previously shown, the human osteosarcoma cell line HOS 58 represents a differentiated stage of osteoblast development. The potential of HOS 58 for still further in vitro differentiation suggests the line can serve as a model of osteoblast maturation. Using this cell line, we have investigated the influence of 1,25-(OH)(2)-D(3) (D(3)), TGF-beta and Dexamethasone (Dex) on proliferation and on the protein and mRNA levels of alkaline phosphatase (AP), procollagen 1 (Col 1), and osteocalcin (Oc), as well as mineralization during 28 days in culture. AP mRNA and protein were highly expressed throughout the culture period with further increase of protein AP activity at constant gene expression levels. A differentiation inhibiting effect of either TGF-beta or Dex was seen. Col 1 was investigated without the use of ascorbic acid and showed only minor changes during culture time or stimulation. The gene expression for Oc increased continually whereas protein synthesis peaked at confluence and decreased thereafter. TGF-beta and Dex treatments decreased Oc mRNA and protein levels. Stimulation by D3 was maximal at day 7 with a decrease thereafter. HOS 58 cells showed no mineralization capacity when stimulated with different agents, as measured by energy-dispersive X-ray microanalysis. This was not due to absence of Cbfa1 expression. In conclusion, the HOS 58 osteosarcoma cell line represents a differentiated cell line with highly expressed and physiologically regulated AP expression during further differentiation in culture. We observed a dissociation between osteocalcin gene expression and protein secretion which may contribute to the lack of mineralization in this cell line.
机译：培养的啮齿动物成骨细胞重申体内观察到的成骨细胞的表型分化和成熟。如先前所示，人骨肉瘤细胞系HOS 58代表成骨细胞发育的分化阶段。 HOS 58在进一步体外分化中的潜力表明，该系可以作为成骨细胞成熟的模型。使用该细胞系，我们研究了1,25-（OH）（2）-D（3）（D（3）），TGF-beta和地塞米松（Dex）对增殖以及蛋白质和mRNA水平的影响培养中28天的碱性磷酸酶（AP），前胶原1（Col 1）和骨钙素（Oc）的矿化程度。在整个培养期间，AP mRNA和蛋白高度表达，在恒定的基因表达水平下，蛋白AP活性进一步提高。观察到TGF-β或Dex的分化抑制作用。在不使用抗坏血酸的情况下对Col 1进行了研究，在培养时间或刺激过程中仅显示了微小变化。 Oc的基因表达持续增加，而蛋白质合成在融合时达到峰值，此后下降。 TGF-β和Dex处理可降低Oc mRNA和蛋白质水平。在第7天，D3刺激最大，此后减少。通过能量色散X射线微分析法测量，当用不同的试剂刺激时，HOS 58细胞没有矿化能力。这不是由于没有Cbfa1表达。总之，HOS 58骨肉瘤细胞系代表分化的细胞系，在进一步分化培养过程中具有高度表达和生理调节的AP表达。我们观察到骨钙素基因表达与蛋白质分泌之间的分离可能导致该细胞系缺乏矿化作用。

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