Basic study on the differentiation induction of human germ cell tumors and its clinical application

人生殖细胞肿瘤分化诱导的基础研究及其临床应用

• 批准号: 01570919
• 负责人: SEKIYA Souei
• 金额: $ 1.34万
• 依托单位: Chiba 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570919/
• 关键词: germ cell tumor; HMBA; N-myc; neuron specific enolase; neural rosette; extracellular matrix; 神経特異エノラ-ゼ

1. A plupribotent human embryonal carcinoma cell line (NEC14) could be induced to differentiate morphologically by treatment with 10^<-2>M N, N'-hexamethylene-bis-acetamide (HMBA) in vitro. The differentiation accompanied the marked induction of human major histocompatibility antigens (HLA-A, B, C), and the changes of stage specific embryonic antigens (from SSEA-1^-/SSEA-3^+ to SSEA-1^+/SSEA-3^-). Differentiated NEC14 cells expressed mesenchym-associated molecules, vimentin and tenascin ; thus HMBA can induce these cells to certain mesenchymalle elements of embryonal mesoderm.2. The level of N-myc expression was high in undifferentiated NEC14 cells, but decreased transiently to less than 1/10 of the original level after the HMBA induction. To investigate the role of N-myc gene in the NEC14 cell differentiation, we established several NEC14 transformants expressing exogenous N-myc genes. They showed shorter population doubling time, increased plating efficiency and tumorigenic potential.3. The level of neuron specific enolase (NSE) in serum was found to be a novel diagnostic marker for immature teratomas and dysgerminomas.4. From the human teratocarcinoma line PA-1, we established a clonal line that produced a distinct neural rosettes within in vitro multicellular spheroids. This line expressed neuroectodermal antigens such as A2B5, HNK-1 and NC-1, NSE, S-100 protein and vimentin. These rosette showed a strikingly polarized and overlapped deposition of extracellular matrix components including laminin, type IV collagen, heparan sulfate proteoglycan and tenascin.

1.多能人胚胎癌细胞系(NEC14)可以通过在体外用10^-2M N，N'-六亚甲基双乙酰胺(HMBA)处理来诱导形态学分化。分化伴随着人类主要组织相容性抗原（HLA-A、B、C）的显着诱导，以及阶段特异性胚胎抗原的变化（从SSEA-1^-/SSEA-3^+到SSEA-1^+/SSEA-3^-）。分化的NEC14细胞表达间充质相关分子、波形蛋白和腱蛋白；因此HMBA可以诱导这些细胞形成胚胎中胚层的某些间充质成分。 2． N-myc 表达水平在未分化的 NEC14 细胞中较高，但在 HMBA 诱导后短暂下降至原始水平的 1/10 以下。为了研究N-myc基因在NEC14细胞分化中的作用，我们建立了几个表达外源N-myc基因的NEC14转化体。它们表现出更短的群体倍增时间、更高的铺板效率和致瘤潜力。3．血清中神经元特异性烯醇化酶(NSE)水平被认为是未成熟畸胎瘤和无性细胞瘤的新诊断标志物。 4．我们从人类畸胎瘤细胞系 PA-1 中建立了一个克隆系，该克隆系在体外多细胞球体中产生了独特的神经玫瑰花结。该细胞系表达神经外胚层抗原，如 A2B5、HNK-1 和 NC-1、NSE、S-100 蛋白和波形蛋白。这些玫瑰花结显示出细胞外基质成分的显着极化和重叠沉积，包括层粘连蛋白、IV型胶原、硫酸乙酰肝素蛋白聚糖和生腱蛋白。

项目成果

Hasegawa,T.et al.: "A transient decrease in Nーmyc expression and its biological role during differentiation of human embryonal carcinoma cells." Differentiation.

Hasekawa, T. 等人：“N-myc 表达的短暂减少及其在人胚胎癌细胞分化过程中的生物学作用。”

Makoto Kawata et al.: "Neural rosette formation within in vitro spheroids of a clonal huiman terots carciusima cell line,PAー1/NR:role of extracellwlar matrix componeuts in the morphogenesis." Cancen Research.

Makoto Kawata 等人：“克隆 huiman terots carciusima 细胞系体外球体内的神经玫瑰花结形成，PA-1/NR：细胞外基质成分在癌症研究中的作用。”

Makoto Kawata: "Neuron-Specific Enolase as a Serum Marker for Immature Teratoma and Dysgerminoma" Gynecologic Oncology. 32. 191-197 (1989)

Makoto Kawata：“神经元特异性烯醇化酶作为未成熟畸胎瘤和无性细胞瘤的血清标志物”妇科肿瘤学。

Sekiya, S., Kimura, H., Yamazawa, K., Kera, K., Kawata, M., Takamizawa. H., and Oda, K.: "Induction of human embryonal carcinoma cell differentiation using N, N'-Hexamethylene Bisacetamide in vitro. Gynecol." Oncol.36. 69-78 (1990)

关谷，S.，木村，H.，山泽，K.，克拉，K.，川田，M.，高见泽。

Souei Sekiya st al.: "Inoluction of Human Embryonal Carcinoma Cell Differentiation Using N,N^´ーHexamethylene Bisacetamide in Vitro" Gynecologic Oncology. 36. 69-78 (1990)

Souei Sekiya 等人：“体外使用 N,N^´ー六亚甲基双乙酰胺注射人类胚胎癌细胞分化”《妇科肿瘤学》36. 69-78 (1990)。