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Expression of Serum Amyloid A Transcripts in Human Bone Tissues Differentiated Osteoblast-Like Stem Cells and Human Osteosarcoma Cell Lines

机译:血清淀粉样蛋白A转录本在人骨组织成骨细胞样干细胞和人骨肉瘤细胞系中的表达

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摘要

Although the liver is the primary site of cytokine-mediated expression of acute-phase serum amyloid A (SAA) protein, extrahepatic production has also been reported. Besides its role in amyloidosis and lipid homeostasis during the acute-phase, SAA has recently been assumed to contribute to bone and cartilage destruction. However, expression of SAA in human osteogenic tissue has not been studied. Therefore, we first show that SAA1 (coding for the major SAA isoform) but not SAA2 transcripts are expressed in human trabecular and cortical bone fractions and bone marrow. Next, we show expression of (i) IL-1, IL-6, and TNF receptor transcripts; (ii) the human homolog of SAA-activating factor-1 (SAF-1, a transcription factor involved in cytokine-mediated induction of SAA genes); and (iii) SAA1/2 transcripts in non-differentiated and, to a higher extent, in osteoblast-like differentiated human mesenchymal stem cells. Third, we provide evidence that human osteoblast-like cells of tumor origin (MG-63 and SAOS-2) express SAF-1 under basal conditions. SAA1/2 transcripts are expressed under basal conditions (SAOS-2) and cytokine-mediatedconditions (MG-63 and SAOS-2). RT-PCR, Western blot analysis, andimmunofluorescence technique confirmed cytokine-mediated expression of SAA onRNA and protein level in osteosarcoma cell lines while SAA4, a protein ofunknown function, is constitutively expressed in all osteogenic tissuesinvestigated.
机译:尽管肝脏是细胞因子介导的急性期血清淀粉样蛋白A(SAA)蛋白表达的主要部位,但也有肝外产生的报道。除了在急性期的淀粉样变性和脂质体内平衡中发挥作用外,SAA最近还被认为有助于破坏骨骼和软骨。但是,尚未研究SAA在人成骨组织中的表达。因此,我们首先表明,SAA1(编码主要的SAA亚型)而不是SAA2转录物在人的小梁和皮质骨部分以及骨髓中表达。接下来,我们显示(i)IL-1,IL-6和TNF受体转录物的表达; (ii)SAA激活因子-1(SAF-1,一种参与细胞因子介导的SAA基因诱导的转录因子)的人类同源物; (iii)SAA1 / 2转录本未分化,并且在更高程度上在成骨细胞样分化的人间充质干细胞中表达。第三,我们提供证据表明,在基础条件下,人类肿瘤来源的成骨细胞样细胞(MG-63和SAOS-2)表达SAF-1。 SAA1 / 2转录本在基础条件下表达(SAOS-2),并由细胞因子介导条件(MG-63和SAOS-2)。 RT-PCR,蛋白质印迹分析和免疫荧光技术证实了细胞因子介导的SAA的表达骨肉瘤细胞系中的RNA和蛋白质水平,而SAA4是一种功能未知,在所有成骨组织中组成性表达调查。

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