Sugimoto T, Kato T, Sawada T, Horii Y, Kemshead J T, Hino T, Morioka H, Hosoi H
Children's Research Hospital, Kyoto, Japan.
Cancer Res. 1988 May 1;48(9):2531-7.
In normal development the neural crest gives rise to sympathetic neuroblasts, sensory and autonomic ganglia, as well as Schwann cells. One tumor arising from this tissue is the neuroblastoma (NB), a malignancy of the adrenergic component of the sympathetic nervous system. Recent histological studies have shown that neuroblastomas can present with a schwannian cell component, rich in S100 protein. We have investigated the differentiation of NB cell lines, GOTO and RT-LN-1, into a schwannian cell phenotype using bromodeoxyuridine (BrdU). This agent induced morphological changes in these cell lines. Flat-epithelial cells were identified in the GOTO cell line and both flat-epithelial and neuronal phenotypes were found in the RT-LN-1 cell line. S100 protein (beta-Subunit) was induced in both cell lines after 18-25 days of BrdU treatment as determined by enzyme-linked immunoassay. In addition increase in the beta-subunit of S100 protein was identified in BrdU-treated flat-epithelial cells by indirect immunofluorescence using a monoclonal antibody specific for the beta-subunit of the protein. Cyclic nucleotide phosphodiesterase activity significantly increased in both BrdU-treated NB cell lines, as compared with nontreated cells. However no significant increase of glial fibrillary acidic protein in BrdU-treated cells was found either by enzyme-linked immunoassay or indirect immunofluorescence using a monoclonal antibody to glial fibrillary acidic protein. Thus, cells with Schwann cell characteristics can clearly be identified in the neuroblastoma cell lines after BrdU treatment. Fluorescence-activated cell sorting analysis revealed no quantitative changes in cell membrane antigens recognized by monoclonal antibodies UJ-13A (neuroectodermal associated antigen) and anti-Thy-1 (Thy-1) on BrdU treatment. In contrast, UJ-127-11 (neuroectodermal associated) decreased, and W6/32 and BB7.7 (HLA-ABC) and BBM.1 (beta 2-microglobulin) markedly increased in both BrdU-treated cell lines. No induction of L243 (HLA-DR), B7/21 (HLA-DP), and Genox 3.55 (HLA-DQ) was noted. The increased HLA-ABC (HLA class I) antigen may enable BrdU-treated NB cells to be recognized by cytotoxic T-cells. This may be related to the pathological evidence that NB patients whose tumors are rich in S100 protein have a better prognosis. Further studies on the potential of differentiation agents to induce a phenotypic change, that is associated with an improved prognosis for NB patients, are required.
在正常发育过程中，神经嵴产生交感神经母细胞、感觉和自主神经节以及施万细胞。源自该组织的一种肿瘤是神经母细胞瘤（NB），它是交感神经系统肾上腺素能成分的恶性肿瘤。最近的组织学研究表明，神经母细胞瘤可呈现富含S100蛋白的施万细胞成分。我们使用溴脱氧尿苷（BrdU）研究了NB细胞系GOTO和RT-LN-1向施万细胞表型的分化。该试剂诱导了这些细胞系的形态变化。在GOTO细胞系中鉴定出扁平上皮细胞，在RT-LN-1细胞系中发现了扁平上皮和神经元两种表型。通过酶联免疫测定法测定，在BrdU处理18 - 25天后，两种细胞系中均诱导产生了S100蛋白（β亚基）。此外，使用针对该蛋白β亚基的单克隆抗体通过间接免疫荧光法在BrdU处理的扁平上皮细胞中鉴定出S100蛋白β亚基增加。与未处理的细胞相比，两种经BrdU处理的NB细胞系中环状核苷酸磷酸二酯酶活性均显著增加。然而，无论是通过酶联免疫测定法还是使用针对胶质纤维酸性蛋白的单克隆抗体进行间接免疫荧光法，在经BrdU处理的细胞中均未发现胶质纤维酸性蛋白有显著增加。因此，在BrdU处理后的神经母细胞瘤细胞系中可以清楚地鉴定出具有施万细胞特征的细胞。荧光激活细胞分选分析显示，在BrdU处理后，单克隆抗体UJ-13A（神经外胚层相关抗原）和抗Thy-1（Thy-1）识别的细胞膜抗原没有定量变化。相反，在两种经BrdU处理的细胞系中，UJ-127-11（神经外胚层相关）减少，而W6/32和BB7.7（HLA-ABC）以及BBM.1（β2微球蛋白）显著增加。未观察到L243（HLA-DR）、B7/21（HLA-DP）和Genox 3.55（HLA-DQ）的诱导。HLA-ABC（I类HLA）抗原的增加可能使经BrdU处理的NB细胞能够被细胞毒性T细胞识别。这可能与病理学证据相关，即肿瘤富含S100蛋白的NB患者预后较好。需要进一步研究分化剂诱导与NB患者预后改善相关的表型变化的潜力。
