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; 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.

已经表明，细胞周期与凋亡相关。细胞周期停滞对细胞凋亡的影响有争议对细胞类型或刺激。以前，我们发现用L-莫Mimosine或KIP 1强制表达KIP 1在G 1期诱导细胞周期停滞，并在包括小鼠B淋巴瘤线WEHI-23 I的各种细胞中的细胞中凋亡，但是到目前为止，细胞周期和凋亡之间连接之间的分子机制尚不清楚。因此，我们试图分离与细胞周期停滞诱导的细胞凋亡有关的基因。我们通过使用逆转录病毒载体从WEHI-23 1细胞或脾细胞生成cDNA库，并引入WEHI-23 I细胞。用L-omosine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片段作为反义方向插入载体。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KIP I诱导的细胞凋亡。此外，C-MYC的过表达增加了KIP 1诱导的细胞凋亡，并通过强制表达MAD4反转了C-MYC的抑制作用。这些结果表明，C-MYC是一个至关重要的分子，它决定细胞周期停滞时细胞的命运。此外，我们获得了抑制细胞周期停滞诱导凋亡的cDNA片段。这些碎片的分析正在进行中。