亚砷酸钠对NE-4C细胞Nrf2及Hippo信号通路的影响

Effects of NaAsO 2 on Nrf2 and Hippo Signal Pathway in NE-4C Cells

目的探讨亚砷酸钠对NE-4C细胞的核因子-类胡萝卜素2相关因子(nf-e2-related factor 2,Nrf2)通路及河马(Hippo)信号通路的影响。方法以不同浓度亚砷酸钠干预细胞48h,采用CCK-8法检测细胞活力;分别采用Western blot法检测亚砷酸钠对NE-4C细胞Nrf2通路相关蛋白的影响,qRT-PCR检测亚砷酸钠对NE-4C细胞Hippo信号通路相关分子mRNA表达的影响。结果亚砷酸钠干预NE-4C细胞48h后,除0.2、0.4μmol/L组外,各干预组细胞活力随亚砷酸钠浓度的增加而降低;亚砷酸钠干预后抗氧化蛋白指标Nrf2、血红素加氧酶1(heme oxygenase-1,HO-1)蛋白表达升高,Kelch样ECH联合蛋白1(Kelch like ECH associated protein 1,Keap1)蛋白表达降低,呈剂量依赖性,提示亚砷酸钠可诱导NE-4C细胞产生氧化应激进而激活抗氧化系统;Hippo信号通路核心分子哺乳动物STE20激酶(mammalian STE20-like protein kinase 1,MST1)、Salvador同系物1(salvador homolog 1,SAV1)、Mps结合激酶激活物1(Mps once binder kinase activator like 1,MOB1)、大肿瘤抑制基因1(large tumor suppressor homolog 1,LATS1)、Yes相关蛋白(Yes-associated protein,YAP)mRNA的表达水平随亚砷酸钠浓度的增加而升高。结论亚砷酸钠可抑制NE-4C细胞增殖,诱导细胞产生氧化应激并激活Hippo信号通路。

Objective To investigate the effects of NaAsO_(2)on nf-e2-related factor 2(Nrf2)pathway and hippo(Hippo)pathway in NE-4C cells.Methods The cells were treated with different concentrations of NaAsO_(2)for 48h,and the cell viability was detected by CCK-8 method.The effects of NaAsO_(2)on Nrf2 pathway-related proteins in NE-4C cells were detected by Western blot,and the effects of NaAsO_(2)on the expression of Hippo signal pathway-related molecules mRNA in NE-4C cells were detected by qRT-PCR.Results After NE-4C cells were treated with NaAsO_(2)for 48h,except for 0.2 and 0.4μmol/L group,the cell viability decreased with the increase of NaAsO_(2)concentration,the expression of antioxidant protein indexes Nrf2 and HO-1 increased and the expression of Keap1 protein decreased in a dose-dependent manner,suggesting that NaAsO_(2)can induce oxidation in NE-4C cells and activate antioxidant system.The expression of MST1,SAV1,MOB1,LATS1 and YAP mRNA,the core molecules of Hippo signaling pathway,increased with the increase of NaAsO_(2)concentration.Conclusion NaAsO_(2)can inhibit the proliferation of NE-4C cells,induce oxidative stress and activate Hippo signal pathway.