尤文氏肉瘤中nm23-H1/NDPK-A的高表达:矛盾的免疫组化反应性及缺乏预后意义
High nm23-H1/NDPK-A expression in Ewing tumors: paradoxical immunohistochemical reactivity and lack of prognostic significance.
作者信息
Aryee D N, Ströbel T, Kos K, Salzer-Kuntschik M, Zoubek A, Veron M, Ambros I M, Traincart F, Gadner H, Kovar H
机构信息
Children's Cancer Research Institute, St Anna Kinderspital, Vienna, Austria.
出版信息
Int J Cancer. 1995 Apr 21;64(2):104-11. doi: 10.1002/ijc.2910640206.
Expression of nm23-H1/NDPK-A has been reported to correlate inversely with metastasizing potential of rodent experimental cells and some human tumors. In the search for reliable molecular prognostic indicators for Ewing tumors (ET), a group of aggressive presumably neuroectodermal malignancies in children and adolescents, we studied nm23-H1/NDPK-A expression. Northern-blot and RT-PCR analyses were employed to semi-quantificatively measure nm23-H1 mRNA levels in ET cell lines and tissue extracts. A panel of monoclonal antibodies (MAbs) were used to evaluate protein abundance by Western blotting and immunohistochemistry. The nm23-H1/NDPK-A gene was also investigated on the DNA level to define possible genomic alterations. Our results revealed neither nm23-H1 allelic loss nor gene amplification and failed to show any significant variation in nm23-H1 mRNA or NDPK-A protein levels of primary or metastatic ET. NDPK-A protein levels were high and comparable to those of MCF-7 breast-cancer cells and to aggressive stage-IV neuroblastoma cell lines. nm23-H2/NDPK-B expression in ET was slightly more variable but generally lower than in MCF-7 cells. In the immunohistochemical analysis, however, discrepancies in the reactivity patterns with different antibodies were observed. Differential sensitivity to various fixation methods and heat treatment pointed to a structurally polymorphic NDPK-A protein. nm23-H1 expression studies using immunohistochemistry for prognostic counselling should thus be interpreted with caution.
据报道,nm23-H1/NDPK-A的表达与啮齿动物实验细胞和一些人类肿瘤的转移潜能呈负相关。在寻找尤因肿瘤(ET)可靠的分子预后指标时,我们研究了nm23-H1/NDPK-A的表达,ET是儿童和青少年中一组具有侵袭性的、推测为神经外胚层来源的恶性肿瘤。采用Northern印迹法和逆转录-聚合酶链反应(RT-PCR)分析对ET细胞系和组织提取物中的nm23-H1 mRNA水平进行半定量测定。使用一组单克隆抗体(MAb)通过蛋白质印迹法和免疫组织化学法评估蛋白质丰度。还在DNA水平上研究了nm23-H1/NDPK-A基因,以确定可能的基因组改变。我们的结果显示,原发性或转移性ET中既没有nm23-H1等位基因缺失,也没有基因扩增,并且nm23-H1 mRNA或NDPK-A蛋白水平也没有任何显著变化。NDPK-A蛋白水平较高,与MCF-7乳腺癌细胞以及侵袭性IV期神经母细胞瘤细胞系相当。ET中nm23-H2/NDPK-B的表达变化略大,但总体上低于MCF-7细胞。然而,在免疫组织化学分析中,观察到不同抗体的反应模式存在差异。对各种固定方法和热处理的不同敏感性表明NDPK-A蛋白存在结构多态性。因此,使用免疫组织化学进行nm23-H1表达研究以提供预后咨询时应谨慎解释。