›› 2011, Vol. 23 ›› Issue (4): 275-278.doi: 10.3969/j.issn.1004-616x.2011.01.001

• 论著 • 上一篇    下一篇

N-甲基-N'-硝基-N-亚硝胍抑制细胞表皮生长因子受体酪氨酸激酶活性

王为民, 高永生   

  1. 中国计量学院生命科学学院药学系,浙江 杭州 310018
  • 收稿日期:2011-03-16 修回日期:2011-04-18 出版日期:2011-07-30 发布日期:2011-07-30
  • 通讯作者: 王为民

N-methyl-N'-nitro-N-nitrosoguanidine suppresses protein tyrosine kinase activity of epidermal growth factor receptor

WANG Wei-min,GAO Yong-sheng   

  1. Department of Pharmacy, College of Life Sciences, China Jiliang University, Hangzhou 310018, Zhejiang, China
  • Received:2011-03-16 Revised:2011-04-18 Online:2011-07-30 Published:2011-07-30
  • Contact: WANG Wei-min

摘要: 目的: 研究低浓度N-甲基-N'-硝基-N-亚硝胍 (MNNG)诱发细胞表皮生长因子受体 (EGFR)形成的二聚体,干扰了EGFR介导的下游信号转导功能的分子机制。 方法:构建EGFR胞内域真核表达的载体,转染Lec-1细胞,纯化EGFR胞内域重组蛋白质后,分别以不同浓度的MNNG (0.25、0.5和1.0 μmol/L MNNG)处理EGFR 1 h,应用酶联免疫吸附测定法检测MNNG各浓度对重组EGFR胞内域蛋白质酪氨酸激酶活性的影响。 结果:MNNG浓度大于0.5 μmol/L即可显著下调EGFR胞内域酪氨酸活性 (P<0.01)。 结论 :MNNG通过抑制EGFR胞内域的酪氨酸激酶活性而阻断其对下游信号的传递。

关键词: 表皮生长因子受体, N-甲基-N' -硝基-N-亚硝胍, 酪氨酸激酶

Abstract: OBJECTIVE: To elucidate the molecular mechanism of low concentration of N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) that interfered with the function of EGFR- mediated cellular signal transduction pathway,by inducing the epidermal growth factor receptor(EGFR)clustering,which was similar to that of epidermal growth factor treatment. METHODS: Eukaryotic expression vector of EGFR cytoplasmic domain gene was constructed and transfected into Lec-1 cells. Then the recombinant EGFR cytoplasmic domain protein was purified,and treated with MNNG for 1 h,at concentrations of 0.25,0.5,and 1.0 μmol/L. Enzyme linked immunoassay was used to measure the protein tyrosine kinase activity of recombinant EGFR cytoplasmic domain. Untreated recombinant cytoplasmic domain protein tyrosine kinase was used as control. RESULTS:Compared with normal control group,tyrosine kinase activity of EGFR was significantly inhibited by 0.5 μmol/L MNNG (P<0.01). CONCLUSION:Low concentration of MNNG interfered with the signal transduction pathway of EGFR by supressing its tyrosine kinase activity.

Key words: epidermal growth factor receptor, N-methyl-N'-nitro-N-nitrosoguanidine, tyrosine kinase