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目的探讨c-Jun N端激酶(JNK)信号通路在As2O3诱导乳腺癌细胞凋亡中的作用。方法体外培养MCF7细胞,以As2O3为刺激源,溴化丙锭(PI)染色结合流式细胞术方法检测细胞周期及细胞凋亡;双荧光素酶报告基因法检测MCF7细胞中AP-1的诱导活化;Western印迹法检测JNK、c-Jun的诱导活化及Fra-1的蛋白表达。结果 As2O3可显著诱导MCF7细胞凋亡并导致细胞周期行进阻滞;As2O3可诱导JNK持续性高强度表达,转录因子AP-1的转录激活活性上调;As2O3可诱导c-Jun活化并诱导Fra-1表达;转染c-Jun显性负性突变体TAM67或Fra-1shRNA可有效降低As2O3诱导的乳腺癌细胞凋亡。结论 JNK/AP-1途径是介导As2O3诱导乳腺癌细胞凋亡反应的重要信号通路。
Objective To investigate the role of c-Jun N-terminal kinase (JNK) signaling pathway in As2O3-induced breast cancer cell apoptosis. Methods MCF7 cells were cultured in vitro, and the cell cycle and apoptosis were detected by using propidium iodide (As2O3) as stimulus and flow cytometry. The induction of AP-1 in MCF7 cells was detected by dual luciferase reporter assay Activation and Western blotting were used to detect the activation of JNK and c-Jun and the protein expression of Fra-1. As2O3 could induce the apoptosis of MCF7 cells and block the cell cycle progression. As2O3 induced the sustained high intensity expression of JNK and the transcriptional activation of transcription factor AP-1. As2O3 induced c-Jun activation and induced the expression of Fra-1 The transfection of the negative mutant of c-Jun, TAM67 or Fra-1 shRNA, can effectively reduce As2O3-induced breast cancer cell apoptosis. Conclusion JNK / AP-1 pathway is an important signal pathway that mediates As2O3-induced apoptosis in breast cancer cells.