氢醌诱导TK6细胞miR-221表达异常致PARP-1基因下调的机制

Hydroquinone Induced Down-regulated Expression of PARP-1 and miR-221 in TK6 cells

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 explore the mechanism of decreased PARP-1 expression induce by hydroquinone.Methods Hydroquinone dissolved with PBS buffer,2.5,5.0 and 10.0 μmol/L hydroquinone were given to the TK6 cells,respectively,and the cells treated with PBS as the control.MiR-221 expression was defined by reverse transcription real-time RT-PCR assay,PARP-1 protein expression was detected by immunoblotting,and potential target gene for miR-221 was analyzed by using bioinformatics methods.Results The expression of PARP-1 protein shown a negative association with the doses of hydroquinone,miR-221 expression were inhibited by 0.17-,0.42-fold by 5.0 and 10.0 μmol/L hydroquinone,respectively.A regulatory element complex was observed between miR-221 and 3＇untranslated region（UTR） of MBD2 mRNA.Conclusion PARP-1 inactivation induced by hydroquinone may be associate with the down-regulated expression of miR-221.