DIFFERENTIATION OF HUMAN GERM-CELL TUMOR-CELLS INVIVO AND INVITRO
DIFFERENTIATION OF HUMAN GERM-CELL TUMOR-CELLS INVIVO AND INVITRO
复制标题
DOI：
10.1267/ahc.25.563
复制
发表时间：
1992-01-01
影响因子：
2.4
通讯作者：
MARUYAMA, T
中科院分区：
文献类型：
--
作者：
HATA, J;FUJIMOTO, J;MARUYAMA, T
从睾丸生殖细胞肿瘤中建立了3株细胞系不同、分化能力不同的人生殖细胞肿瘤细胞系。用维甲酸（RA）诱导对应于卵黄囊瘤的NCR-G1产生甲胎蛋白（AFP）。NCR-G2和NCR-G3是人胚胎癌（EC）细胞系。它们具有人类EC细胞所共有的细胞学和免疫表型特征，如在其他EC细胞中所描述的。NCR-G2在培养基中以漂浮细胞聚集体的形式生长，而NCR-G3以漂浮细胞聚集体的形式生长，并使细胞贴附于培养皿表面。NCR-G3的这些扁平细胞以化学方式表达多种分化抗原，包括肌源性标志物、胚外滋养层细胞标志物（人绒毛膜促性腺激素（hCG））和细胞外蛋白。当用RA处理时，NCR-G3细胞的分化能力变得更加明显。流式细胞仪分析表明，人EC细胞表面标记2D 7、2 H2和5D4，即我们从NCR-G2细胞免疫获得的小鼠单克隆抗体，从RA处理的NCR-G3细胞的细胞表面消失。与这些发现一致，附着在培养皿上的NCR-G3扁平细胞对这些抗体呈阴性。此外，较长时间暴露于RA增强hCG的生产，暴露时间依赖性的方式。这些观察结果清楚地表明，NCR-G3具有多能分化能力，并且它们分化成滋养层细胞谱系而不是体细胞。与NCR-G3相反，NCR-G2在RA处理下未显示任何分化能力。所有三种生殖细胞肿瘤系在无胸腺小鼠中均以100%的效率产生肿瘤。在NCR-G1荷瘤小鼠的血清中观察到高AFP含量。NCR-G3荷瘤小鼠血清中检测到高水平的hCG和AFP。因此，我们新建立的人生殖细胞肿瘤细胞系为人类胚胎发生和EC细胞分化的分子机制提供了新的见解。
Three in vitro human germ cell tumor cell lines with different cell lineage and variable capability of differentiation were established from testicular germ cell tumors. NCR-G1 corresponding to yolk sac tumor were induced a-fetoprotein (AFP) production with retinoic acid (RA) treatment. NCR-G2 and -G3 were human embryonal carcinoma (EC) cell lines. They have cytological and immuno-phenotypic characteristics common to human EC cells as has been described in other EC cells. NCR-G2 grew as floating cell aggregates in the culture medium, whereas NCR-G3 grew as floating cell aggregate and flattered cells attached to the surface of culture dish. These flattened cells of NCR-G3 immunohistochemically expressed a variety of differentiation antigens including myogenic markers, extraembryonic trophoblastic cell marker (human chorionic gonadotropin (hCG)), and extracellular proteins. Differentiation capabilities of NCR-G3 cells became more evident when they were treated with RA. By flowcytometric analysis, human EC cell surface markers 2D7, 2H2 and 5D4 which were mouse monoclonal antibodies we obtained from immunization by NCR-G2 cells, disappeared from cell surface of RA-treated NCR-G3 cells. Consistent with these findings, the flattened cells of NCR-G3 that attached to the culture dish were negative for these antibodies. Moreover, longer exposure to RA enhanced hCG production in exposure time dependent fashion. These observations clearly indicate that NCR-G3 possess multipotent differentiation capabilities and they differentiated into the trophoblastic cell lineage other than somatic cells. In contrast to NCR-G3, NCR-G2 did not show any differentiation capabilities with RA treatment. All three germ cell tumor lines produced tumors in athymic mice with 100% efficiency. High AFP content was observed in the sera of NCR-G1 tumor-bearing mice. High hCG and AFP contents were detected in the sera of the NCR-G3 tumor-bearing mice. Thus, our newly established human germ cell tumor cell lines add new insights of molecular mechanism of human embryogenesis and differentiation of EC cells.