Identification and characterization of genes responsive to apoptosis: application of DNA chip technology and mRNA differential display.
Identification and characterization of genes responsive to apoptosis: application of DNA chip technology and mRNA differential display.
复制标题
细胞凋亡响应基因的鉴定和表征：DNA芯片技术和mRNA差异显示的应用。
DOI：
10.14670/hh-15.1271
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发表时间：
2000
影响因子：
2
通讯作者：
Y. Sun
中科院分区：
文献类型：
--
作者：
Y. Sun
细胞凋亡（程序性细胞死亡）是一种遗传程序性的主动细胞死亡过程，用于维持生理条件下的稳态和响应各种刺激。许多人类疾病与细胞凋亡增加（如AIDS和神经退行性疾病）或细胞凋亡减少（如癌症和自身免疫性疾病）相关。为了了解细胞凋亡信号通路和凋亡相关基因，我们建立了两种细胞模型系统，分别用DNA损伤剂依托泊苷和氧化还原剂1，10-菲咯啉（OP）诱导细胞凋亡。利用DNA芯片分析或mRNA差异显示（DD）来鉴定对这两种药物诱导的细胞凋亡有反应的基因。在依托泊苷与芯片杂交模型中，我们定义了以p53依赖性方式（通过激活p53靶基因如Waf-1/p21、PCNA、GPX、S100 A2和PTGF-β）以及以p53非依赖性方式（通过激活ODC和TGF-β受体等）介导凋亡的信号通路。在DD筛选的OP模型中，我们克隆并鉴定了两个基因：谷胱甘肽合成酶，编码参与谷胱甘肽合成的酶和凋亡敏感基因（SAG），一个新的进化保守基因，编码锌环指蛋白。这两种基因似乎都能保护细胞免受氧化还原剂诱导的细胞凋亡。对SAG的进一步研究表明，它是酵母中的一个生长必需基因，属于一个新发现的促进蛋白质泛素化和降解的基因家族。通过这种活性，SAG调节细胞周期进程和许多其他关键的生物学过程。因此，SAG可能是抗癌和抗炎治疗的有效药物靶标。
Apoptosis (programmed cell death) is a genetically programmed active cell death process for maintaining homeostasis under physiological conditions and for responding to various stimuli. Many human diseases have been associated with either increased apoptosis (such as AIDS and neurodegenerative disorders) or decreased apoptosis (such as cancer and autoimmune disorders). In an attempt to understand apoptosis signaling pathway and genes associated with apoptosis, we established two cell model systems on which apoptosis is induced either by DNA damaging agent, etoposide or by redox agent, 1,10-phenanthroline (OP). DNA chip profiling or mRNA differential display (DD) was utilized to identify genes responsive to apoptosis induced by these two agents. In etoposide model with chip hybridization, we defined signaling pathways that mediate apoptosis in p53 dependent manner (through activation of p53 target genes such as Waf-1/p21, PCNA, GPX, S100A2 and PTGF-beta) as well as in p53-independent manner (through activation of ODC and TGF-beta receptor, among others). In OP model with DD screening, we cloned and characterized two genes: glutathione synthetase, encoding an enzyme involved in glutathione synthesis and Sensitive to Apoptosis Gene (SAG), a novel evolutionarily conserved gene encoding a zinc RING finger protein. Both genes appear to protect cells from apoptosis induced by redox agents. Further characterization of SAG revealed that it is a growth essential gene in yeast and belongs to a newly identified gene family that promotes protein ubiquitination and degradation. Through this activity, SAG regulates cell cycle progression and many other key biological processes. Thus, SAG could be a valid drug target for anti-cancer and anti-inflammation therapies.