Cytopathological studies of Ewing's sarcoma and an application of pathologic diagnosis using newly produced monoclonal antibodies

1. 本文建立了3株人尤文氏肉瘤细胞系，对其形态学、细胞遗传学特征和分子分析进行了研究。染色体分析显示三种细胞系中有两种发生易位，t(11; 22)(q24; q12)，与先前报道的Ewing肉瘤和骨骼和软组织的原始神经外胚层肿瘤相同。细胞培养物的形态不受dbcAMP的影响。神经元特异性烯醇化酶在培养细胞、培养液上清液和裸鼠血清中含量较高。利用经典杂交瘤技术制备了两个单克隆抗seki抗体。这些靶抗原在尤文氏肉瘤、神经母细胞瘤、横纹肌肉瘤、恶性淋巴瘤等癌的肿瘤细胞中均有表达。N-myc、C-myc、Nras、ets-1和PDGFB癌基因的southern blot和Northern blot分析结果对尤文氏肉瘤没有特异性。我们通过形态学、免疫组化和细胞遗传学研究将尤文氏肉瘤与横纹肌肉瘤和小细胞骨肉瘤进行了比较。神经母细胞瘤的形态学特征在光镜和免疫组织化学上很容易与尤文氏肉瘤混淆。我们建立了一种新的神经母细胞瘤细胞系（HNB- 18），并研究了神经母细胞瘤与尤文氏肉瘤的区别。上述结果（1-6）提示尤文氏肉瘤可能起源于原始神经外胚层细胞。虽然从我们的工作来看，尤文氏肉瘤和原始神经外胚层肿瘤之间的差异尚不清楚，但我们认为这两种肿瘤可能是通过组织学分化程度来表达的。

1. Three human Ewing's sarcoma lines were established, and these were investigated for their morphological and cytogenetic characterization and molecular analysis.2. Chromosome analysis revealed a translocation, t(11 ; 22)(q24 ; q12)in two of three lines, as reported previously for Ewing's sarcoma and primitive neuroectodermal tumors of bones and soft tissues.3. Morphology of cell cultures was unaffected by exposure to dbcAMP. Values of neuron-specific enolase were high in cultured cells, supematant of the culture fluid, and nude mice serum.4. Two clones of monoclonal anti-Seki antibodies were produced using the classical hybridoma technology. Expression of these target antigens was found in tumor cells of Ewing's sarcoma, neuroblastoma and osteosarcoma among tumor cells of Ewing's sarcoma, neuroblastoma, osteosarcoma rhabdomyosarcoma, malignant lymphoma and other carcinomas examined.5. The findings of Southem blot and Northern blot analyses of the N-myc, C-myc, Nras, ets-1 and PDGFB oncogenes were not specific for Ewing's sarcoma.6. We compared Ewing's sarcoma with rbabdomyosarcoma and small cell osteosarcoma by morphological, immunobistochemical and cytogenetic studies. The morphological features of neuroblastoma is easily confused with Ewing's sarcoma by light microscopy and immunohistochemistry. We established a new neuroblastoma cell line(HNB- 18)and investigated a difference between neuroblastoma and Ewing's sarcoma. These results described above(1-6)suggest that Ewing's sarcoma may originate in a primitive neuroectodermal cell. Although a difference between Ewing's sarcoma and primitive neuroectodermal tumor is still unclear from our work, we consider that these two tumors may be expressed by the histological grade of differentiation.