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目的探讨氢醌(HQ)致TK6细胞PARP-1基因表达下调的机制。方法以磷酸盐缓冲液(PBS)溶解HQ,以PBS处理组为对照组,分别以2.5、5.0和10.0μmol/L HQ染毒TK6细胞为染毒组。应用实时荧光定量-聚合酶链反应检测miR-221的表达改变,以蛋白印迹法检测PARP-1蛋白表达量,以生物信息学方法分析miR-221的潜在甲基化相关靶基因。结果与对照组相比,2.5、5.0和10.0μmol/L HQ组细胞PARP-1蛋白量逐渐下降,5.0和10.0μmol/L HQ组miR-221表达量分别下降17%(P<0.05)、42%(P<0.05)。miR-221能与MBD2 mRNA的3’非翻译区(UTR)形成调控复合体。结论 HQ致PARP-1基因下调机制可能与miR-221表达异常有关。
Objective To investigate the mechanism of hydroquinone (HQ) down-regulation of PARP-1 gene expression in TK6 cells. Methods HQ was dissolved in phosphate buffered saline (PBS) and treated with PBS. The cells were treated with 2.5, 5.0 and 10.0 μmol / L HQ, respectively. The expression of miR-221 was detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). The expression of PARP-1 protein was detected by Western blotting. The potential methylation-related target genes of miR-221 were analyzed by bioinformatics methods. Results Compared with the control group, the protein levels of PARP-1 in 2.5, 5.0 and 10.0 μmol / L HQ groups decreased gradually, while the expression levels of miR-221 in the 5.0 and 10.0 μmol / L HQ groups decreased by 17% (P <0.05), 42 % (P <0.05). miR-221 forms a regulatory complex with the 3 ’untranslated region (UTR) of MBD2 mRNA. Conclusion The down-regulation mechanism of PARP-1 gene in HQ may be related to the abnormal expression of miR-221.