Beresford J N, Taylor G T, Triffitt J T
Nuffield Department of Orthopaedic Surgery, University of Oxford, England.
Eur J Biochem. 1990 Oct 24;193(2):589-97. doi: 10.1111/j.1432-1033.1990.tb19377.x.
Recombinant human interferon-alpha 2C and recombinant human interferon-gamma (5-1000 U/ml) inhibit the proliferation of normal human bone-derived cells and a human osteosarcoma cell line. In the bone-derived cells the inhibitory effect of interferon-gamma was significantly greater than that of interferon-alpha, whereas in the osteosarcoma cell line the inhibitory effects of both interferons were quantitatively similar. Interferon-alpha did not affect the alkaline phosphatase activity of either type of cells. In contrast, interferon-gamma affected the activity of the enzyme in both cell types: in the bone-derived cells the effect of interferon-gamma was stimulatory whereas in the osteosarcoma cells the effect was inhibitory. In both cell types interferon-gamma selectively inhibited the incorporation of radiolabelled proline into type I collagen. In the osteosarcoma cells, the effects of both interferons on collagen synthesis were quantitatively similar. In the bone-derived cells, however, interferon-alpha decreased proline incorporation into collagen and non-collagen proteins to a similar extent and thus did not affect collagen synthesis when expressed as a percentage of total protein synthesis. Two-dimensional polyacrylamide gel electrophoresis of the radiolabelled proteins of the cell layer synthesised by both cell types in the presence of either interferon demonstrated that this treatment enhanced or induced the synthesis of a total of 21 individual proteins (19 in bone cells, 14 in osteosarcoma), ranging in apparent molecular mass over 14-87 kDa. The set of proteins induced was different in all four combinations of cells and interferon. A tentative identification of several of the proteins was possible based upon estimation of molecular mass, preferential induction by interferon-alpha or interferon-gamma and differential induction in normal and transformed bone-derived cells. The results of this study demonstrate that interferons have complex effects upon the proliferative and biosynthetic activities of human bone-derived cells and demonstrate significant differences between the responses of normal cells and transformed bone-derived cell line. Further investigations will be required in order to determine whether or not these differences are unique to the osteosarcoma cell line or are a characteristic of the effects of interferons on bone-derived cells in general.
重组人干扰素α2C和重组人干扰素γ（5 - 1000 U/ml）可抑制正常人骨源性细胞和人骨肉瘤细胞系的增殖。在骨源性细胞中，干扰素γ的抑制作用明显强于干扰素α，而在骨肉瘤细胞系中，两种干扰素的抑制作用在数量上相似。干扰素α对两种细胞类型的碱性磷酸酶活性均无影响。相比之下，干扰素γ影响两种细胞类型中该酶的活性：在骨源性细胞中，干扰素γ的作用是刺激性的，而在骨肉瘤细胞中，其作用是抑制性的。在两种细胞类型中，干扰素γ均选择性抑制放射性标记的脯氨酸掺入I型胶原蛋白。在骨肉瘤细胞中，两种干扰素对胶原蛋白合成的影响在数量上相似。然而，在骨源性细胞中，干扰素α使脯氨酸掺入胶原蛋白和非胶原蛋白的程度相似，因此当以总蛋白质合成的百分比表示时，并不影响胶原蛋白合成。在存在任一干扰素的情况下，对两种细胞类型合成的细胞层放射性标记蛋白质进行二维聚丙烯酰胺凝胶电泳显示，这种处理增强或诱导了总共21种个体蛋白质的合成（骨细胞中19种，骨肉瘤中14种），表观分子量范围为14 - 87 kDa。在细胞和干扰素的所有四种组合中，诱导产生的蛋白质组各不相同。基于分子量估计、干扰素α或干扰素γ的优先诱导以及正常和转化的骨源性细胞中的差异诱导，对其中几种蛋白质进行了初步鉴定。本研究结果表明，干扰素对人骨源性细胞的增殖和生物合成活性具有复杂影响，并表明正常细胞和转化的骨源性细胞系的反应存在显著差异。为了确定这些差异是否是骨肉瘤细胞系所特有的，还是干扰素对骨源性细胞一般作用的特征，还需要进一步研究。
