SPECIFIC HIGH-AFFINITY BINDING AND BIOLOGIC ACTION OF RETINOIC ACID IN HUMAN NEURO-BLASTOMA CELL-LINES
SPECIFIC HIGH-AFFINITY BINDING AND BIOLOGIC ACTION OF RETINOIC ACID IN HUMAN NEURO-BLASTOMA CELL-LINES
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DOI：
10.1073/pnas.80.18.5525
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发表时间：
1983-01-01
期刊：
影响因子：
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通讯作者：
MANGELSDORF, D
中科院分区：
文献类型：
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作者：
HAUSSLER, M;SIDELL, N;MANGELSDORF, D
神经母细胞瘤细胞是一个很好的神经元发育模型，因为它们有能力延长神经突以响应各种刺激，包括维甲酸。在本实验中，5个人类神经母细胞瘤细胞系（LA-N-1, IMR-32, LA-N-5， SK-N-MC和CHP-100）检测了细胞维甲酸结合蛋白（CRABP）的存在，这是一种与维生素a分子功能有关的受体样蛋白。CRABP通过蔗糖梯度离心，汞试剂对氯苯磺酸（PCMBS）的选择性抑制和饱和度分析进行了鉴定和定量。5个品系均含有显著水平的胞质CRABP （2.5 ~ 7.5 pmol/mg蛋白），具有特异性高亲和维甲酸结合、沉淀系数为2s、PCMBS抑制的特点。其中3个品系（LA-N-1、IMR-32和LA-N-5）的生长受到1 .mu的强烈抑制。而2 （LA-N-1和La-N-5）则明显分化为星状、梭状形态，并伴有典型的神经突生长。在这些条件下，SK-N-MC和CHP-100系对维甲酸的抗增殖作用具有相对的抗性。然而，这5个品系在软琼脂中形成不依赖于锚定的结肠的能力都受到视黄酸的有效抑制。因此，尽管CRABP在单层培养中不一定与生长抑制有关，但它与维甲酸在软琼脂中抑制神经母细胞瘤集落形成的能力有关。需要更多的实验来确定这种对软琼脂生长的影响是否反映了维甲酸将致瘤性神经母细胞瘤细胞系转化为正常分化表型的假定能力。
Neuroblastoma cells are a good model for neuronal development because of their ability to extend neurites in response to various stimuli, including retinoic acid. In the present experiments, 5 human neuroblastoma cell lines (LA-N-1, IMR-32, LA-N-5, SK-N-MC, and CHP-100) were examined for the presence of cellular retinoic acid binding protein (CRABP), a receptor-like protein implicated in the molecular functioning of vitamin A. CRABP is identified and quantitated by sucrose gradient centrifugation, selective inhibition by the mercurial reagent p-chloromercuribenzene sulfonic acid (PCMBS), and saturation analysis. All 5 lines contain significant levels of cytosolic CRABP (2.5-7.5 pmol/mg of protein), which dispaly typical properties of specific high affinity retinoic acid binding, a sedimentation coefficient of 2 S, and inhibition by PCMBS. Three of the lines (LA-N-1, IMR-32, and LA-N-5) are strongly growth inhibited by 1 .mu.M retinoic acid in monolayer culture, whereas 2 (LA-N-1 and La-N-5) undergo marked differentiation to a stellate, fusiform morphology with characteristic neurite outgrowths. The SK-N-MC and CHP-100 lines are relatively resistant to the antiproliferative effects of retinoic acid under these conditions. Nevertheless, all 5 lines are effectively inhibited by retinoic acid in their ability to form anchorage-independnet colones in soft agar. Thus, although CRABP is not necessarily correlated with growth inhibition in monolayer culture, it is associated with retinoic acid''s ability to inhibit neuroblastoma colony formation in soft agar. More experiments will be required to determine if this effect on growth in soft agar reflects the putative ability to retinoic acid to convert tumorigenic neuroblastoma cell lines into the normal differentiated phenotype.