14,15-EET通过EGFR和p44/42 MAPK信号通道诱导肿瘤细胞增殖(英文)  

14,15-Epoxyeicosatrienoic Acid Induces Proliferation of Tumor Cells via EGFR and p44/42 MAPK Signaling Pathways

作　　者：程黎明[1] 赵硕生[1] 王勇[1] 李旭光[1] 汪道文[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院内科学系和基因治疗中心,武汉市430030

出　　处：《医学分子生物学杂志》2007年第5期406-409,共4页Journal of Medical Molecular Biology

摘　　要：目的探讨14,15-EET促进肿瘤细胞增殖的机制。方法Tca-8113培养于含10%小牛血清的DMEM,必要时更换无血清DMEM使其进入静止期。用14,15-EET和/或抑制剂处理后收集细胞蛋白,用Western印迹检测EGFR和p44/42MAPK(ERK1/ERK2)的磷酸化水平。结果14,15-EET以剂量依赖性方式刺激EGFR和p44/42MAPK(ERK1/ERK2)蛋白磷酸化,该效应能够被特异性的EGFR抑制剂AG1478阻断。MTT分析显示,AG1478能够完全取消14,15-EET诱导的Tca-8113细胞增殖效应。结论EGFR的活化是花生四烯酸细胞色素P450表氧化酶代谢产物14,15-EET丝裂原信号途径中的关键事件;在该途径中EGFR的活化是p44/42MAPK(ERK1/ERK2)活化的上游事件。Objective To explore the mechanism of how 14, 15-epoxyeicosatrienoic acids （EET） stimulates tumor cell proliferation. Methods Human tongue squamous cell carcinoma de- rived Tca-8113 cells were cultured in DMEM containing 10 % bovine serum and, when necessary, made quiescent by replacing with serum-free DMEM. After treated with 14, 15-EET or inhibitors, cellular proteins were collected, and the phosphorylation of EGFR and p44/42 MAPK （ ERK1/ ERK2） was detected by western blot assay. Results It was shown that 14, 15-EET stimulates tyrosine phosphorylation of both EGFR and p44/42 MAPK （ERK1/ERK2） in a concentration-de- pendent fashion. Meanwhile, this EET-induced phosphorylation was blocked by EGFR-specific in- hibitor AG1478, also in a dose-dependent manner. The analysis of 3- （4, 5-dimethylthiazol-2- yl） -2, 5-diphenyltetrazolium bromide （MTI＇） revealed that AG1478 completely abolished the mi- togenic induction by 14, 15-EET in Tca-8113 cells. Conclusion The results suggest that EGFR activation is a crucial event in mitogenic signaling transmission of the P450 arachidonate epoxygenase metabolite 14, 15-EET, and exerts an upstream regulatory effect on p44/42 MAPK （ERK1/ERK2） activation in EET-mediated signaling pathway.

关 键 词：14 15-环氧化二十碳三烯甘油酸 EGFR P44/42 MAPK(ERK1/ERK2) 细胞增殖 

分 类 号：R73[医药卫生—肿瘤]