CAJ | 학술논문

目的：观察氢醌对HL-60细胞向单核细胞分化的影响，并初步探讨miR-146a的调控作用。方法：设miR-146a-5p抑制剂和阴性对照处理的HL-60细胞，分别以不同浓度(0、0.5、1.0、2.5和5.0μmol/L)的氢醌处理3 h后，接种于含豆蔻酰佛波醇(PMA)的培养基中，培养72 h诱导其分化。采用实时荧光定量PCR检测细胞内miR-146a及其靶基因TRAF6的表达量，流式细胞术检测CD11b和CD14的表达，Western blot检测TRAF6蛋白及其下游调控因子IκBα蛋白的表达水平。结果：与对照组相比(0μmol/L)，氢醌可抑制PMA诱导的HL-60细胞CD11b和CD14的表达，5.0μmol/L的氢醌使其表达量分别减少44%和38%(P均<0.01)；同时miR-146a的表达比对照组升高近2倍(P<0.01)；TRAF6 mRNA的表达下降约40%，其蛋白表达下降74%(P均<0.01)；磷酸化IκBα蛋白的表达减少约45%，IκBα总蛋白表达升高约73%(P均<0.01)。当抑制miR-146a的表达后，氢醌对TRAF6、IκBα和磷酸化IκBα蛋白表达的影响不明显。结论：miRNA-146a是氢醌影响HL-60细胞分化的调控因子之一。
목적：관찰경곤대HL-60세포향단핵세포분화적영향，병초보탐토miR-146a적조공작용。방법：설miR-146a-5p억제제화음성대조처리적HL-60세포，분별이불동농도(0、0.5、1.0、2.5화5.0μmol/L)적경곤처리3 h후，접충우함두구선불파순(PMA)적배양기중，배양72 h유도기분화。채용실시형광정량PCR검측세포내miR-146a급기파기인TRAF6적표체량，류식세포술검측CD11b화CD14적표체，Western blot검측TRAF6단백급기하유조공인자IκBα단백적표체수평。결과：여대조조상비(0μmol/L)，경곤가억제PMA유도적HL-60세포CD11b화CD14적표체，5.0μmol/L적경곤사기표체량분별감소44%화38%(P균<0.01)；동시miR-146a적표체비대조조승고근2배(P<0.01)；TRAF6 mRNA적표체하강약40%，기단백표체하강74%(P균<0.01)；린산화IκBα단백적표체감소약45%，IκBα총단백표체승고약73%(P균<0.01)。당억제miR-146a적표체후，경곤대TRAF6、IκBα화린산화IκBα단백표체적영향불명현。결론：miRNA-146a시경곤영향HL-60세포분화적조공인자지일。
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 5.0μmol/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.