Molecular mechanism of cell cycle arrest-induced apoptosis

细胞周期阻滞诱导细胞凋亡的分子机制

• 批准号: 14599001
• 负责人: ADACHI Takahiro
• 金额: $ 1.98万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14599001/
• 关键词: apoptosis; cell cycle; kip1; cyclin dependent kinase inhibitor; Kip1

It has been shown that cell cycle has relevance to apoptosis. Effect of cell cycle arrest on apoptosis is controversial upon cell types or stimuli. Previously we found that treatment with L-mimosine or enforced expression of Kip 1 induced cell cycle arrest at G 1 phase and apoptosis in various types of cells including mouse B lymphoma line WEHI-23 I. But so far, molecular mechanism of the connection between cell cycle and apoptosis remains unclear. Therefore, we tried to isolate genes related to cell cycle arrest-induced apoptosis. We generated that cDNA library from WEHI-23 1 cells or, spleen cells by using retrovirus vectors and introduced into WEHI-23 I cells. Cells were treated with L-mimosine and integrated cDNA fragments were recovered from the live cells by PCR. We obtained three independent partial c-Myc cDNA fragments were inserted into the vector as antisense direction. Expression of these constructs in WEHI-23 1 cells repressed c-Myc expression and inhibited Kip i-induced apoptosis. Moreover, overexpression of c-Myc augmented Kip 1-induced apoptosis and inhibition of c-Myc by enforced expression of Mad4 reversed it. These results indicated that c-Myc is a crucial molecule which decide cell fate upon cell cycle arrest. Furthermore, we obtained cDNA fragments that inhibit cell cycle arrest-induced apoptosis. Analyses of these fragments are in progress.

研究表明，细胞周期与细胞凋亡有关。细胞周期停滞对细胞凋亡的影响因细胞类型或刺激而存在争议。此前我们发现L-含羞草碱处理或强制表达Kip 1可诱导包括小鼠B淋巴瘤系WEHI-23 I在内的多种类型细胞的细胞周期停滞在G 1期并发生凋亡。但迄今为止，两者之间联系的分子机制尚不明确。细胞周期和细胞凋亡仍不清楚。因此，我们尝试分离与细胞周期停滞诱导的细胞凋亡相关的基因。我们使用逆转录病毒载体从 WEHI-23 1 细胞或脾细胞中生成 cDNA 文库，并将其引入 WEHI-23 I 细胞中。用L-含羞草碱处理细胞并通过PCR从活细胞中回收整合的cDNA片段。我们获得了三个独立的部分c-Myc cDNA片段，以反义方向插入到载体中。这些构建体在 WEHI-23 1 细胞中的表达抑制了 c-Myc 表达并抑制了 Kip i 诱导的细胞凋亡。此外，c-Myc 的过表达会增强 Kip 1 诱导的细胞凋亡，而通过强制表达 Mad4 来抑制 c-Myc 则可逆转这种情况。这些结果表明c-Myc是细胞周期停滞时决定细胞命运的关键分子。此外，我们获得了抑制细胞周期停滞诱导的细胞凋亡的cDNA片段。对这些片段的分析正在进行中。

项目成果

Hirai, H., Adaclli, T., Tsubata, T.: "Involvement of cell cycle progression in survaival signaling through CD40 in Blymphocyte line WEHI-231"Cell Death Differ. 11. 261-269 (2004)

Hirai, H.、Adaclli, T.、Tsubata, T.：“B 淋巴细胞系 WEHI-231 中通过 CD40 参与细胞周期进程的生存信号传导”细胞死亡有所不同。

Hokazono, Y., Adachi, T., Wabl, M., Tada, N., Amagasa, T., Tsubata, T.: "Inhibitory co-receptors activated by antigens but not by anti-immunoglobulin heavy chain antibodies install requirement of co-stimulation through CD40 for survival and proliferation

Hokazono, Y.、Adachi, T.、Wabl, M.、Tada, N.、Amagasa, T.、Tsubata, T.：“抑制性共受体由抗原激活，但不由抗免疫球蛋白重链抗体激活，安装要求

Hirai, H., Adachi, T., Tsubata, T.: "Involvement of cell cycle progression in survaival signaling through CD4O in B lymphocyte line WEHI-231."Cell Death Differ.. 11. 261-269 (2004)

Hirai, H.、Adachi, T.、Tsubata, T.：“B 淋巴细胞系 WEHI-231 中通过 CD4O 参与细胞周期进程的生存信号传导。”细胞死亡差异.. 11. 261-269 (2004)

C.Wakabayashi, T.Adachi, J.Wienands, T.Tsubata: "A distinct signaling pathway used by the IgG-containing B cell antigen receptor"Science. 298. 2392-2395 (2002)

C.Wakabayashi、T.Adachi、J.Wienands、T.Tsubata：“包含 IgG 的 B 细胞抗原受体使用的独特信号传导途径”《科学》。

Hokazono, Y., Adachi, T., Wabi, M., Tada, N., Amagasa, T., Tsubata, T.: "Inhibitory co-receptors activated by antigens but not by anti-inununoglobulin heavy chain antibodies install requirement of co-stimulation through CD4O for survival and proliferation

Hokazono, Y.、Adachi, T.、Wabi, M.、Tada, N.、Amagasa, T.、Tsubata, T.：“抑制性共受体由抗原激活，但不由抗免疫球蛋白重链抗体激活，安装要求