氢醌抑制HL-60细胞分化上调过氧化物酶2-Cys过氧化物氧还蛋白表达

Hydroquinone inhibits HL-60 cells differentiation and upregulates 2-Cys peroxiredoxins expression

目的研究氢醌（HQ）对HL-60细胞向单核系、粒系分化的影响。方法 HQ 1,5和50μmo·lL^-1分别与豆蔻酰佛波醇乙酯（PMA）20 nmol·L^-1或1.25%DMSO共同处理细胞,分别于48和96 h收集细胞。通过观察细胞形态、硝基四氮唑蓝还原反应鉴定细胞分化;CCK-8检测细胞增殖,荧光定量PCR检测2-Cys过氧化物氧还蛋白（Prxs）基因表达的变化;Western印迹检测2-Cys Prxs蛋白表达的变化。结果在HQ1-50μmol·L^-1作用下,HL-60细胞向单核系分化受到抑制;HQ1和5μmol·L^-1对DMSO诱导的粒系分化无影响,但HQ50μmol·L^-1可抑制细胞向粒系分化。HQ5和50μmol·L^-1与诱导剂的共同作用可以抑制HL-60细胞增殖。与正常对照组比较,PMA和DMSO组2-Cys Prxs基因表达水平均有降低的趋势,HQ1,5和50μmol.L^-1＋PMA20 nmol·L^-1组PrxⅠ,PrxⅢ和PrxⅣ各个基因表达水平与PMA组比较均有增高的趋势;DMSO诱导分化组中仅HQ 50μmol·L^-1＋1.25%DMSO组PrxⅠ和PrxⅢ基因表达水平与单独DMSO组比较显著增高（P〈0.05）。结论 HQ1和5μmol.L^-1可以抑制HL-60细胞向单核系分化,HQ50μmol·L^-1可抑制细胞向粒系分化,并上调PrxⅠ和PrxⅢ基因的表达。

OBJECTIVE To explore the effect of hydroquinone（HQ） on HL-60 cells differentiation to monocytes or granulocytes.METHODS HL-60 cells were cotreated with HQ 1,5 and 50 μmol·L^-1 and phorbol 12-myristate 13-acetate（PMA） 20 nmol·L^-1 or 1.25%DMSO.They were collected after cultured for 48 and 96 h,respectively.Differentiation of monocytes and granulocytes was assessed by Giemsa stain and the reduction of nitroblue tetrazolium assay（NBT）.Cell proliferation was detected by CCK-8.Real-time PCR and Western blotting were performed to examine the expression of 2-Cys peroxiredoxins（2-Cys Prxs）.RESULTS HQ 1-50 μmol·L^-1 inhibited monocytic differentiation induced by PMA,HQ 1 and 5 μmol·L^-1 had no effect on differentiation to granulocytes induced by DMSO,but HQ 50 μmol·L^-1 could inhibit differentiation to granulocytes.HQ 5 and 50 μmol·L^-1 and combined with the inducer could inhibit cell proliferation.The expression of 2-Cys Prxs decreased on HL-60 during monocytic or granulocytic differentiation induced by PMA or DMSO.Compared with PMA 20 nmol·L^-1 group,the expression of PrxⅠ,PrxⅢ and PrxⅣ in HQ different doses combined with PMA 20 nmol·L^-1 groups had tended to increase.However,for granulocytic differentiation,only HQ 50 μmol·L^-1 increased the expression of PrxⅠ and PrxⅢ significantly compared with DMSO control group（P〈0.05）.CONCLUSION HQ 1 and 5 μmol·L^-1 inhibit HL-60 cells differentiation to monocytes,while HQ 50 μmol·L^-1 inhibits differentiation to granulocytes and up-regulates the expression of PrxⅠ and PrxⅢ.