Pedersen W A, Becker L E, Yeger H
Department of Pathology, Hospital for Sick Children, Toronto, Ontario, Canada.
Int J Cancer. 1993 Feb 1;53(3):463-70. doi: 10.1002/ijc.2910530319.
A series of human neuroblastoma (NB) cell lines was analyzed for expression of peripherin, a class-III intermediate filament protein expressed at high levels in ganglia of the peripheral nervous system. By Western blotting, peripherin protein was detected in all human NB cell lines examined. The highest level of peripherin was found in the NUB-7 cell line, previously characterized as homogeneously neuroblastic. By immunofluorescence labeling, peripherin was shown to be organized in a perinuclear filamentous pattern and, exemplified by IMR32 cells, was also shown to be localized to spontaneously formed neurites. Peripherin was expressed in neuroblastic but not substrate-adherent cells, and was found at low levels in I-type cells. There was a pronounced redistribution of peripherin to neurites formed in response to dibutyryl cyclic adenosine monophosphate (dbcAMP) and all-trans-retinoic acid (RA). In NUB-7 cells, which do not extend neurites in response to nerve growth factor, there was no change in the level of peripherin protein following treatment with this agent. Both dbcAMP and RA induced a redistribution of peripherin to neurite extensions, but only treatment with RA increased the level of the protein as demonstrated with NUB-6A4 and NUB-6C4 subclones. Peripherin was also variably expressed in peripheral neuroepithelioma (NE) cell lines tested, but was organized into a more basket-like filamentous pattern in these cells. The heterogeneous expression and distribution of peripherin in NB and NE cell lines indicate that this protein is associated with maturation of the neuronal phenotype and hence serves as a differentiation marker for tumors derived from the neural crest.
对一系列人类神经母细胞瘤（NB）细胞系进行了分析，以检测外周蛋白的表达情况。外周蛋白是一种III类中间丝蛋白，在外周神经系统的神经节中高水平表达。通过蛋白质免疫印迹法，在所检测的所有人类NB细胞系中均检测到了外周蛋白。在外周蛋白水平最高的NUB-7细胞系中，该细胞系先前被鉴定为均一性成神经细胞。通过免疫荧光标记显示，外周蛋白以核周丝状模式排列，以IMR32细胞为例，还显示其定位于自发形成的神经突。外周蛋白在成神经细胞中表达，但在贴壁细胞中不表达，在I型细胞中表达水平较低。在二丁酰环磷酸腺苷（dbcAMP）和全反式维甲酸（RA）诱导形成的神经突中，外周蛋白有明显的重新分布。在对神经生长因子无反应而不延伸神经突的NUB-7细胞中，用该因子处理后外周蛋白水平没有变化。dbcAMP和RA均诱导外周蛋白重新分布到神经突延伸部位，但只有用RA处理才能增加蛋白水平，如NUB-6A4和NUB-6C4亚克隆所示。在所检测的外周神经上皮瘤（NE）细胞系中，外周蛋白也有不同程度的表达，但在这些细胞中其排列成更像篮状的丝状模式。外周蛋白在NB和NE细胞系中的异质性表达和分布表明，该蛋白与神经元表型的成熟有关，因此可作为神经嵴来源肿瘤的分化标志物。
