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

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:20μmol/L AOH作用NIH3T3细胞1 h,或用PKA特异性抑制剂10μmol/L H89预处理1 h,再进行20μmol/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.

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 subunits of PKA in NIH3T3 cells were observed by immunofluorescence 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.