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Tumorigenicity and Analysis of the Cell Growth of Human Medulloblastoma

人髓母细胞瘤的致瘤性和细胞生长分析

基本信息

批准号：
01570809
负责人：
SHIMIZU Keiji
金额：
$ 1.34万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

Medulloblastoma are among the most common malignant brain tumors in children. It suggested that the precursors of these tumors consisted of immature bipotential cells which could differentiate into neuronal and glial cells. We established two human medulloblastoma cell lines in order to investigate their characterization and cell differentiation. One was derived from a 2-year-old girl with a cerebellar tumor (designated as ONS-76 cells), and another was from a 9-year-old girl with a metastatic tumor in the right lower frontal lobe from her posterior fossa (designated as ONS-81 cells). The in vitro population-doubling times were 18.6 and 19.2 hours, respectively. These cells were transplantable into the nude mice subcutaneously and intrathecally. Experimental models with meningeal dissemination were produced by intracisternal inoculation of ONS-76 cells into the nude mice. All mice were dead within 65 days after 1x107 tumor cells were inoculated intracisternally. Their median survival t … More ime (MST) was 56 days.Immunohistochemical studies showed that both cells possessed neurofilament protein (NFP) (Mr 145 K and 200K) and neuron-specific enolase (NSE), without glial fibrillar acidic protein (GFAP) or S-100 protein. These cell lines also had epidermal growth factor (EGF) receptor (R), Transferrin-R, and lnsulinーR. These cells could grow in the growth medium without fetal calf serum (FCS). Most these cells possessed class I major histocompatibility complex (MHC) antigens, and these class I antigens were able to be enhanced by human interferon-g (IFN-g). Both cell lines didn't have class II MHC antigen, but these antigens could be induced by human IFN-g. ONS-76 and ONS-81 cells expressed proto-oncogene (N-myc and c-src) products. These cells showed drug-induced differentiation under 50-100 mM of dibutyryl cyclic-adenosine 3', 5'-monophosphate (dbt-cAMP). Fluorescence-activated cell sorter (FACScan) analysis also showed that these both cell lines increased expression of pp60^<c-src> products and NFP (145 K and 200 K). A monoclonal antibody (mAb) (ONS-M-1.0) of lgG2 isotype was raised against medulloblastoma by immunization of mice with ONS--76 cells. This mAb bound these two human medulloblastoma (ONS-76 and -81) cells selectively. Natural killer cells were demonstrated a little ONS-M-1.0 antigens. Biochemical analysis showed that these antigens were sensitive to trypsin but not to neuraminidase, and that they were heat-stable. The high specificity of this antibody^* on medullobalstoma cells might be contributory to the diagnostic and therapeutic strategies for medulloblastoma. Less

髓母细胞瘤是儿童最常见的恶性脑肿瘤之一。这表明这些肿瘤的前体由未成熟的双能细胞组成，可以分化为神经元和神经胶质细胞。我们建立了两种人髓母细胞瘤细胞系，以研究它们的特征和细胞分化。一个来自一名患有小脑肿瘤的 2 岁女孩（指定为 ONS-76 细胞），另一个来自一名 9 岁女孩，该女孩的后颅窝右下额叶有转移性肿瘤（指定为 ONS-81 细胞）。体外群体倍增时间分别为18.6和19.2小时。这些细胞可通过皮下和鞘内移植到裸鼠体内。通过将ONS-76细胞接种到裸鼠脑池内建立脑膜播散实验模型。脑池内接种1x107个肿瘤细胞后，所有小鼠均在65天内死亡。它们的中位生存期 (MST) 为 56 天。免疫组织化学研究表明，这两种细胞都含有神经丝蛋白 (NFP)（Mr 145 K 和 200K）和神经元特异性烯醇化酶 (NSE)，不含胶质纤维酸性蛋白 (GFAP) 或 S-100 蛋白。这些细胞系还具有表皮生长因子 (EGF) 受体 (R)、转铁蛋白-R 和胰岛素 R。这些细胞可以在不含胎牛血清（FCS）的生长培养基中生长。大多数这些细胞具有 I 类主要组织相容性复合体 (MHC) 抗原，并且这些 I 类抗原能够被人干扰素-g (IFN-g) 增强。两种细胞系均不具有 II 类 MHC 抗原，但这些抗原可以被人 IFN-g 诱导。 ONS-76 和 ONS-81 细胞表达原癌基因（N-myc 和 c-src）产物。这些细胞在 50-100 mM 二丁酰环腺苷 3', 5'-单磷酸 (dbt-cAMP) 下表现出药物诱导的分化。荧光激活细胞分选仪 (FACScan) 分析还表明，这两种细胞系均增加了 pp60^<c-src> 产物和 NFP（145 K 和 200 K）的表达。通过用 ONS--76 细胞免疫小鼠，产生抗髓母细胞瘤的 IgG2 同种型单克隆抗体 (mAb) (ONS-M-1.0)。该 mAb 选择性结合这两种人髓母细胞瘤（ONS-76 和 -81）细胞。自然杀伤细胞被证明有少量ONS-M-1.0抗原。生化分析表明这些抗原对胰蛋白酶敏感但对神经氨酸酶不敏感，并且它们是热稳定的。该抗体对髓母细胞瘤细胞的高度特异性可能有助于髓母细胞瘤的诊断和治疗策略。较少的