増殖抑制誘導能をもつ糖転移酵素遺伝子の解析

具有诱导生长抑制能力的糖基转移酶基因分析

• 批准号: 09254268
• 负责人: 辻 崇一
• 金额: $ 1.41万
• 依托单位: The Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09254268/
• 关键词: ガングリオシド; 増殖抑制

最近、本研究者等は、神経芽腫瘍細胞(Neuro2aのサブクローン)にGD3合成酵素のcDNAを導入したところ、細胞はGD3とb-系列ガングリオシドを発現するようになり、増殖が抑制されることを明らかにした。本年度は、この現象をさらに解析し、以下の結果を得た。1.人為的発現制御系を用いた増殖抑制過程の解析:テトラサイクリン応答系を組み込むことにより、GD3合成酵素遺伝子発現から増殖が抑制されるまで経時的な解析が再現性良く行えることが明らかとなった。テトラサイクリン処理後8時間後にはGD3合成酵素mRNAの発現が見られ、以後ほぼ定常的にその発現が見られた。合成されてくるガングリオシドを分析した結果、まずGD3の発現がみられ、以後次第によりシアル酸を多く持つb-系列ガングリオシドの発現が見られた。この間に、細胞の増殖は次第に抑制され、最終的には増殖が停止した。2.増殖抑制過程で発現してくる遺伝子のクローニングとその解析:上記増殖抑制過程で新たに発現あるいは不活化される遺伝子をDifferential display法を用いて検索を行っている。現在のところほぼ80%の検索が完了した。今までに、52の異なるDNA断片の新たな発現もしくは発現増大が見られた。さらにRNA protection analysisを行いfalse positiveの排除などを進めた結果、このなかで10の異なる遺伝子が新たに発現あるいは発現増大してくることが明らかになった。

Recently, our researchers have demonstrated that the cDNA for GD3 synthase was introduced into neuroblastoma cells (Neuro2a) and that the expression of GD3 and b-series genes was inhibited by the gene. This year, the phenomenon was analyzed, and the following results were obtained. 1. The analysis of artificial reproduction control system in the process of proliferation inhibition: GD3 synthase gene production control system is composed of two groups, GD3 synthase gene production control system and GD3 synthase gene production control system. 8 hours after treatment, the expression of GD3 synthase mRNA was observed, and the expression of GD3 synthase mRNA was observed after treatment. The results of the analysis showed that GD3 was found to be the most important factor in the synthesis of GD3, and the b-series was found to be the most important factor in the synthesis of GD3. The growth of these cells is gradually inhibited, and finally stopped. 2. The process of growth inhibition is developed, and the gene is activated. The Differential display method is applied to the process of growth inhibition. 80% of the search is now complete. Now, 52 new DNA fragments have been discovered, and the number of new DNA fragments has increased. The RNA protection analysis was performed in the middle of the false positive exclusion process. The results showed that the RNA protection analysis was performed in the middle of the false positive elimination process. The RNA protection analysis was performed in the middle of the false positive elimination process. The RNA protection analysis was performed in the middle of the false positive elimination process. The RNA protection analysis was performed in the middle of the false positive elimination process.

项目成果

Hong Liu: "Regulated expression system for GD3 synthase cDNA and induction of differentiation in Neuro2a cells." Glycobiology. 7・8. 1067-1076 (1997)

Hong Liu：“GD3 合酶 cDNA 的调节表达系统和 Neuro2a 细胞分化的诱导。” 7・8 (1997)。