N-甲基-N′-硝基-N-亚硝基胍诱发的vero细胞蛋白酪氨酸磷酸化改变

Alteration of protein tyrosine phosphorylation in vero cells induced by N-methyl-N′-nitro-N-nitrosoguanidine

为了研究细胞信号转导通路在致突变物 N-甲基 - N′-硝基 - N-亚硝基胍 ( MNNG)致非定标性突变作用中的地位 ,猴肾 vero细胞在 MNNG处理后 0 ,6,1 2 h制备全细胞抽提液 ,用 Western印迹法观察细胞蛋白质酪氨酸磷酸化的变化 .结果发现 0 .2μmol· L-1MNNG处理 2 .5h后再经 0 h和 1 2 h,45ku蛋白质酪氨酸磷酸化程度增强 ,经 6h时还有62 ku蛋白质酪氨酸磷酸化程度增强 ,1 mg·L-1蛋白合成抑制剂环己米特 ( CHM)处理 vero细胞 1 2 h也可引起 45ku蛋白质酪氨酸磷酸化程度增强 .提示 MNNG处理可能诱发应激相关信号转导通路的激活 .

In order to study the role of cell signal transduction pathways in nontargeted mutagenesis induced by mutagen N methyl N′ nitro N nitrosoguanidine (MNNG), Western blot analysis was used to detect the alteration of protein tyrosine phosphorylation of whole cell extracts of monkey kidney vero cells prepared at 0,6,12 h after 0.2 μmol·L -1 MNNG treatment for 2.5 h. Results showed an increase in degree of tyrosine phosphorylation of 45 ku protein at 0 and 12 h after MNNG treatment,and of 62 ku protein at 6 h after MNNG treatment. Cycloheximide(CHM) treatment (1 mg·L -1 ,12 h) could also increase the degree of tyrosine phosphorylation of 45 ku protein. The results obtained suggest that MNNG induce the activation of stress related signal transduction pathways.