Carcinogenesis, Teratogenesis & Mutagenesis

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The regulatory role of  miR-146a in the differentiation of HL-60 cells inhibited by hydroquinone

YUAN Wei-xin1,JIANG Yan-ping2,XIE Chun-jiao1,QIN Fei1,CHEN Yun-cong1,WEI Qing1,ZENG Xiao-wen1,SHA Yan3,XIAO Yong-mei1,*   

  1. School of Public Health, Sun-Yat Sen University, Guangzhou 510080
  • Received:2014-03-20 Revised:2014-04-04 Online:2014-05-30 Published:2014-05-30

Abstract:

OBJECTIVE: To study the effects of hydroquinone(HQ) on monocytic differentiation of HL-60 cells and explore the regulatory role of miR-146a in the mechanism of toxicity. METHODS: Using the model of monocytic differentiation induction with phorbol 12-myristate 13-acetate (PMA),HL-60 cells were transfected with miR-146a-5p inhibitor or negative control and then treated with various doses of hydroquinone(0,0.5,1.0,2.5 and 5.0 μmol/L) for 3 h,and cells collected after induction with PMA for 72 h. The expressions of miR-146a and its target gene TRAF6,were measured by real-time quantitative PCR. CD11b and CD14 levels were also determined by flow cytometry. Western blot was used to detect protein expression levels of TRAF6,and its downstream regulatory factor IκBα. RESULTS: Hydroquinone could inhibit the expressions of CD11b and CD14 in HL-60 cells induced by PMA. Compared with control group(0 μmol/L),5.0 μmol/L of hydroquinone decreased the expressions of CD11b and CD14 by 44% and 38%,respectively (both P <0.01);the expression of miR-146a increased nearly 2-fold (P<0.01). The expressions of TRAF6 mRNA and protein were decreased by 40% and 74%(P<0.01),respectively. The expression of phosphorylated IκBα protein was reduced by approximately 45%,and the total IκBα protein increased nearly 73% (P<0.01). After inhibiting the expression of miR-146a,there were no significant effects of hydroquinone on TRAF6,phosphorylated IκBα and total IκBα protein expression. CONCLUSION: miRNA-146a may be one of the regulatory factors in which hydroquinone influence HL-60 cells differentiation.

Key words: hydroquinone, HL-60 cells, cell differentiation, microRNA-146a, TRAF6, NF-κB