Isolation, identification and their regulation of genes responsible for the expression of gangliosides.

负责神经节苷脂表达的基因的分离、鉴定及其调控。

• 批准号: 62580111
• 负责人: SANAI Yutaka
• 金额: $ 1.09万
• 依托单位: Department of Biochemistry, Faculty of Medicine, University of Tokyo
• 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62580111/
• 关键词: Ganglioside; oncogene; シアル酸転移酵素; 遺伝子クローニング; ヒトメラノーマ

In order to elucidate the molecular mechanism on the expression of GD3 ganglioside associated with oncogenesis of cells, I examined the isolation of the genomic DNA controlling the expression of GD3 ganglioside which was well known as human melanoma associated antigen by employing cosmid shuttle vector and monoclonal antibody. Transfection of the cosmid library derived from human melanoma cells and repeated cell sortings of the fluorescence-bright population enabled to enrich the antigen positive transfectants. Six groups of indipendent clones were rescued from the GD3 positive transfectants by in vitro packaging procedure. However, these isolated clones lacked the capability of the induction of GD3 in the recipient L cells. These suggested the expression of GD3 in L cells were controlled by more than single gene. Nuclear type oncogenes such as adenovirus E1A and myc were also able to induce GD3 in rat 3Y1 cells, but other type oncogenes (src gene fammily and ras) could induce sialosylparagloboside altematively. These suggested expression of GD3 in transformed cells were related to the common function of the nuclear type oncogene.

为了阐明与细胞肿瘤发生相关的GD3神经节苷脂表达的分子机制，我使用粘粒穿梭载体和单克隆抗体检查了控制GD3神经节苷脂表达的基因组DNA的分离，GD3神经节苷脂是众所周知的人类黑色素瘤相关抗原。源自人黑色素瘤细胞的粘粒文库的转染和荧光亮群体的重复细胞分选能够富集抗原阳性转染子。通过体外包装程序从GD3阳性转染子中拯救出六组独立克隆。然而，这些分离的克隆缺乏在受体L细胞中诱导GD3的能力。这些表明L细胞中GD3的表达受多个基因控制。核型癌基因如腺病毒E1A和myc也能够在大鼠3Y1细胞中诱导GD3，但其他类型癌基因（src基因家族和ras）也可以诱导唾液酸基副红细胞苷。这些表明转化细胞中GD3的表达与核型癌基因的共同功能有关。

项目成果

Y.Sanai,;M.Yamasaki,;Y.Nagai.: Biochim.Biophys.Acta. 958. 368-374 (1988)

Y.Sanai，；M.Yamasaki，；Y.Nagai.：Biochim.Biophys.Acta。

H.Nakaishi,; Y.Sanak,; M.Shibuya,; M.Iwamori,; Y.Nagai.: Cancer Research. 48. 1753-1758 (1988)

H.中石，；

H. Nakaishi: Cancer Res. (1988)

H. Nakaishi：癌症研究。

Y.Sanai,; M.Yamasaki,; Y.Nagai.: Biochm.Biophys.Acta. 958. 368-374 (1988)

Y.Sanai，；

H. Nakaishi; Y. Sanai; M. Shibuya; M. Iwamori; Y. Nagai: "Neosyntheses of neolacto- and novel ganglio-series gangliosides in a rat fibroblastic cell line brought about by transfection with the v-fes oncogene containing Gardner-Arhstein strain feline sarco

H.中石；