国家重点基础研究发展规划项目(973)“鼻咽癌发生发展的基础研究”(G1998051201)和“恶性肿瘤侵袭和转移的机理及分子阻遏”(2002CB513100).
This work was supported by grants from The Special funds for Major State Basic Research of China(G1998051201) and (2002CB513100).
为了探讨EB病毒编码的潜伏膜蛋白1(LMP1)对核转录因子Ets-1表达和活化的影响,并证实细胞外信号调节激酶1/2(ERK1/2)参与了该过程,选用可调控表达LMP1的鼻咽癌细胞系L7,应用蛋白质印迹法检测Ets-1、p-ERK蛋白质表达,免疫共沉淀-蛋白质印迹法检测Ets-1磷酸化状态,使用ERK1/2特异性小分子阻断物PD98059作用后,蛋白质印迹法检测p-ERK、Ets-1表达及磷酸化变化.结果显示:在L7细胞中诱导性LMP1可促进p-ERK、Ets-1蛋白质表达及其苏氨酸残基磷酸化,在一定范围呈时间和剂量效应;通过PD98059对诱导性LMP1作用的干预发现,p-ERK大部分表达被阻断,而Ets-1表达及其苏氨酸磷酸化也被部分阻断,以上结果提示ERK部分介导了LMP1诱导Ets-1表达和活化.
To confirm the effect of EBV encoded LMP1 on expression and activation of nuclear transcription factor, Ets-1 and extracellualr signal regulated kinase(ERK) involved in the process. The expression of p-ERK and Ets-1 were assayed with Western blot, and phosphorylation of Ets-1 was assayed with co-immunoprecipitation-Western blot. MEK1/ERK specific inhibitor PD98059 was used to confirm that ERK mediated the activation of Ets-1 by LMP1. The results showed that in nasopharyngeal carcinoma cell line, EBV-LMP1 enhanced the expression of Ets-1 and p-ERK, and phosphorylation of Ets-1 to some extent in time- and dose-dependent manner. With blockade of PD98059, expression of p-ERK induced by LMP1 decreased significantly, and expression and phosphorylation of Ets-1 by LMP1 decreased partly. These results suggest that expression and phosphorylation of Ets-1 by LMP1 was mediated partly by ERK.
曾亮,李敏,宋鑫,陶永光,唐敏,李嵬,曹亚. EB病毒潜伏膜蛋白1在鼻咽癌细胞中通过ERK介导Ets-1表达[J].生物化学与生物物理进展,2003,30(4):574-578
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