摘要
细胞信号传导PI3K-Akt路径涉及细胞周期、再生、分化、衰老与凋亡的调控,采用高通量生物技术分析基因表达谱可用于抗肿瘤或抗衰老的基因与药物筛选。化学诱变剂甲磺酸乙酯(ethlmethane sulformate,EMS)处理CHO细胞,筛选到抗10μM、20μM浓度lovastatin的E10、E20突变细胞系,将E10细胞再经诱变剂EMS处理,筛选到抗70μM浓度lovastatin的ZE70细胞系。Igf-2刺激基因突变的肌肉萎缩小鼠肌肉细胞系,Western免疫杂交试验结果显示,PI3K-Akt路径的Akt蛋白质磷酸化功能正常。生长因子Igf-2刺激肌肉细胞系C2C12的cDNA基因芯片实验表明,促细胞再生、分化相关的基因表达上调,细胞凋亡相关的基因表达下调。细胞的基因突变,激素、Igf-2等生长因子刺激,往往是肿瘤发生的病因;但是,Igf-2等可用于抗肌肉萎缩疾病,以及抗细胞凋亡与衰老的药物筛选。
Signal Transduetion pathway PI3k is involved in the regulation of cell cycle, proliferation, differentiation, ageing and ap- optosis. Analysis of PI3K pathway by using high through bioteehnology, is benefit for the discovery of anti-cancer or anti-ageing drug. Resuits Showed that Treatment CHO cells by EMS（ethlmethane snlformate） , cell lines of EIO, E20 surviving in 10 μM,20μM lovastatin were selected. Treatment EIO cells by EMS again, a cell line ZE70 surviving in 70 μM lovastatin was isolated. Igf-2 treatment C2C12 and cells isolated from muscular dystrophic mice, the phospho-pAkt was identified well function in both cell lines, therefore, eDNA chips were used for analysis of anti-cancer and anti-apoptosis genes expression in respond to Igf-2 stimulation, and several useful genes were identified
出处
《生物技术通报》
CAS
CSCD
北大核心
2009年第8期134-138,共5页
Biotechnology Bulletin
