互隔交链孢酚对NIH3T3细胞PKA-CREB信号通路的影响
Effect of alternariol on PKA-CREB signal pathway in NIH3T3 cells
马俊芬 1杨璇 2赵继敏 1黄幼田 1杨洪艳 1郑智敏 1董子明1
作者信息
- 1. 郑州大学基础医学院病理生理教研室,河南,郑州,450052
- 2. 郑州大学基础医学院微牛物与免疫教研室,河南,郑州,450052
- 折叠
摘要
目的:探讨互隔交链孢酚(Alternariol,AOH)对小鼠胚胎成纤维细胞NIH3T3中PKA-CREB信号通路的影响.方法:20 μmol/L AOH作用NIH3T3细胞1 h,或用PKA特异性抑制剂10 μmol/L H89预处理1 h,再进行20 μmol/L AOH刺激实验,同时设溶剂对照组.用免疫荧光和免疫细胞化学染色法检测磷酸化PKA催化亚基表达情况,用免疫细胞化学染色法检测磷酸化CREB表达水平.结果:与溶剂对照组相比,AOH诱导NIH3T3细胞中磷酸化PKA催化亚基和磷酸化CREB水平明显增加(P<0.05);用PKA特异性抑制剂H89预处理,降低了AOH激活的PKA催化亚基和CREB的磷酸化水平.结论:AOH能够激活NIH3T3细胞中PKA-CREB信号转导通路,这为探讨AOH致癌的分子机制提供依据.
Abstract
AIM: To investigate the activation of PKA-CREB pathway in NIH3T3 cells induced by AOH. METHODS: The NIH3T3 cells were treated with 20 μmol/L AOH for 1 h, or the cells were pretreated with an inhibitor of PKA ( H89) for 1 h, then exposed to AOH for 1 h. The level of activated catalytic sub-units of PKA in NIH3T3 cells were observed by immunofluores-cence and immunocytochemistry. The phosphorylation of CREB were determined by immunocytochemistry. RESULTS: The expression of phospho-PKA catalytic subunits and phospho-CREB in NIH3T3 cells induced by AOH were significantly higher than that in the control groups ( P < 0. 05 ). H89 partly blocked the AOH-induced phospho-PKA and phospho-CREB expression. CONCLUSION: AOH can induce the activation of PKA-CREB signal pathway in NIH3T3 cells, which provides evidence for the molecular mechanisms of carcinogenesis of AOH.
关键词
互隔交链孢酚/蛋白激酶A/cAMP反应元件结合蛋白/NIH3T3细胞Key words
alternariol/PKA/CREB/NIH3T3/cells引用本文复制引用
基金项目
国家自然科学基金(30471952)
河南省教育厅自然科学基金(2007310020)
出版年
2009
NETL
NSTL
维普
万方数据