【摘要】 目的:探讨互隔交链孢酚(AOH)诱导小鼠胚胎成纤维细胞NIH3T3中DNA聚合酶β(DNA polβ)表达增高的分子信号通路。方法:①15μmol/L AOH作用NIH3T3细胞16 h,利用Western blotting方法检测细胞中DNA polβ的表达,并利用磷酸化CREB抗体检测AOH作用后NIH3T3细胞中CREB信号分子是否被激活。②利用PKA-CREB信号通路特异性抑制剂H89预处理NIH3T3细胞1 h,再加入AOH作用,分别检测磷酸化CREB和DNA polβ表达的变化。结果:①与溶剂对照组相比,AOH诱导NIH3T3细胞中DNA polβ表达明显增高,而且磷酸化CREB蛋白含量也明显增加,差异均显著。②用PKA特异性抑制剂H89预处理,可部分抑制NIH3T3细胞中CREB蛋白的磷酸化,并且降低了AOH引起的NIH3T3细胞中DNA polβ的表达增加。结论:PKA-CREB信号通路在AOH诱导的NIH3T3细胞中DNA polβ表达起重要作用。更多还原

【Abstract】 AIM: To investigate the signal transduction induced DNA polymerase β expression by alternariol(AOH) in NIH3T3 cells.METHODS: The NIH3T3 cells were treated with 15 μmol/L AOH for 16 h or the cells were pretreated with an inhibitor of PKA(H89) for 1 h,and then exposed to AOH.The changes of DNA polβ expression and phospho-CREB in the cells with different treatments were determined by Western blotting and immunocytochemistry analysis.RESULTS: The expression of DNA polymerase β and phospho-CREB in NIH3T3 cells induced by AOH were significantly higher than that in control group(P<0.05).However,H89 partly blocked the AOH-induced phospho-CREB and DNA polβ expression.CONCLUSION: Alternariol up-regulates DNA polymerase β expression via the PKA-CREB pathway in NIH3T3 cells.更多还原