论文部分内容阅读
目的探讨微小RNA 155(miR-155)、BTB和CNC同源蛋白1(BACH1)、醌氧化还原酶1(NQO1)和血红素氧合酶-1(HO-1)在三氧化二砷(ATO)诱导细胞死亡过程中的变化规律,以及miR-155和BACH1可能的调控关系,为增敏ATO治疗新靶点的发现奠定实验基础。方法以不同浓度ATO处理肺腺癌A549细胞后,采用MTT实验检测细胞的存活率或死亡率,试剂盒检测细胞的总抗氧化能力,Westom blot检测BACH1、NQO1和HO-1蛋白的表达,实时荧光定量PCR(qRT-PCR)检测miR-155的表达水平。miR-155类似物(mimic)及其阴性对照转染对数期A549细胞,并设未转染组为对照,qRT-PCR检测miR-155表达水平后,以20μmol/L ATO处理24h,再进行MTT及Western blot检测。结果 10~100μmol/L的ATO可降低细胞的存活率;与空白对照组相比,10、20μmol/L的ATO可减弱细胞的总抗氧化能力,激活BACH1蛋白的表达,抑制miR-155以及NQO1和HO-1蛋白的表达;在转染miR-155mimic后,20μmol/L ATO处理后的A549细胞死亡率低于未转染组和转染阴性对照组,且ATO对BACH1蛋白的激活作用减弱,NQO1和HO-1蛋白表达则高于后两组(P<0.05)。结论ATO可通过抑制miR-155的表达,激活BACH1蛋白,抑制NQO1和HO-1蛋白的表达,从而削弱细胞的总抗氧化能力,最终诱导细胞死亡,提示miR-155和BACH1可作为ATO治疗肺癌过程中的增敏靶点。
Objective To investigate the expression of miR-155, BACH1, NQO1 and HO-1 in arsenic trioxide (ATO) -induced cells The changes in the process of death and possible regulation of miR-155 and BACH1 lay the experimental foundation for the discovery of new targets of hyperthermic ATO therapy. Methods A549 lung adenocarcinoma cells were treated with different concentrations of ATO. MTT assay was used to detect the cell viability or mortality. The kit was used to detect the total anti-oxidative capacity of cells. Western blot was used to detect the expression of BACH1, NQO1 and HO-1. The expression of miR-155 was detected by qRT-PCR. The A549 cells were transfected with miR-155 mimic and its negative control, and untransfected cells were transfected with miR-155. The expression of miR-155 was detected by qRT-PCR and treated with 20 μmol / L ATO for 24 h MTT and Western blot detection. Results ATO of 10-100μmol / L decreased the survival rate of cells. Compared with the blank control group, ATO at 10 and 20μmol / L attenuated the total antioxidant capacity, activated the expression of BACH1 protein, inhibited the expression of miR-155 and NQO1 After transfection with miR-155mimic, the apoptosis rate of A549 cells treated with 20μmol / L ATO was lower than that of untransfected and transfected negative control groups, and the activation of BACH1 protein was attenuated by ATO, NQO1 and HO-1 protein expression was higher than the latter two groups (P <0.05). CONCLUSION ATO inhibits the expression of miR-155, activates BACH1 protein and inhibits the expression of NQO1 and HO-1 proteins, thereby attenuating the total antioxidant capacity of cells and ultimately inducing cell death. It suggests that miR-155 and BACH1 can be used as ATO in the treatment of lung cancer Sensitization target in the process.