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Induction of differentiation on the differentiation of human embryonal carcinoma

诱导分化对人胚胎癌分化的影响

基本信息

批准号：
62570743
负责人：
SEKIYA Souei
金额：
$ 1.22万
依托单位：
Chiba University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

项目摘要

To establish an in vitro differentiation system of human germ cell tumor cells, we initially selected a cell line, NEC 14 from 9 cell lines derived from various types of human germ cell tumors. The NEC 14 line established from testicular embryonal carcinoma ( EC ) showed spontaneously a capacity for multiple differentiation in xenograft tumors in nude mice, such as glandular components, cartilagious tissue, and an extraembryonic syncytiotrophoblastic element. As a differentiation inducing reagent in vitro, we used 10 M of N,N'- hexamethylene bisacetamide ( HMBA ), since this reagent showed the most potent activity to induce apparently differentiated colonies among HMBA, retinoic acid, progesterone, DMSO, and dbc AMP.FACS analyses revealed that the HMBA treatment reduced human EC antigen, SSEA-3, and inversely induced SSEA-1 and HLA-A, B, C antigen on NEC 14 cells. Furthermore, these HMBA-treated NEC 14 cells showed higher adhesivity to plastic substrate, and contained larger amount ( 3X ) of plasminogen activator than untreated NEC 14 cells. After prolonged culture, keratin, desmin, and vimentin were immunohistochemically identifiable on a subpopulation of these HMBA-treated cells.In additional projects, we characterized the molecular properties of F9 embryoglycan recognized by a unique antibody in sera from patients with germ cell tumors. We also found that neuron-specific enolase ( NSE ) measurements are of diagnostic value not only for immature teratomas but also for dysgerminomas.

为了建立人生殖细胞肿瘤细胞的体外分化体系，我们从9个不同类型的人生殖细胞肿瘤细胞系中初步选择了一个细胞系NEC 14。从睾丸胚胎癌（EC）中建立的NEC 14细胞系在裸鼠异种移植肿瘤中自发地表现出多种分化的能力，如腺成分、软骨组织和胚胎外合胞滋养细胞成分。作为体外分化诱导试剂，我们使用了10 M的N，N'-六亚甲基双乙酰胺（HMBA），因为该试剂在HMBA、维甲酸、黄体酮、DMSO和dbc amp之间诱导明显分化菌落的活性最强。facs分析显示，HMBA处理降低了人EC抗原SSEA-3，并在NEC 14细胞上反向诱导SSEA-1和HLA-A、B、C抗原。此外，这些经hmba处理的NEC 14细胞对塑料底物具有更高的粘附性，并且比未经处理的NEC 14细胞含有更多的纤溶酶原激活剂（3倍）。经过长时间的培养，角蛋白、去蛋白和静脉蛋白在这些经hba处理的细胞亚群中被免疫组织化学鉴定出来。在其他项目中，我们在生殖细胞肿瘤患者的血清中鉴定了一种独特的抗体识别F9胚胎蛋白的分子特性。我们还发现，神经元特异性烯醇化酶（NSE）测量不仅对未成熟畸胎瘤，而且对生殖细胞异常瘤也有诊断价值。