miR-146a对THP-1细胞靶基因AUF1调控及细胞因子表达的影响

miR-146a targets at AUF1 and regulates cytokine expression in THP-1 cell line

目的探讨miR-146a对人急性单核白血病细胞系(THP-1)靶基因AU碱基富集区结合降解因子1(AUF1)表达的调控及细胞因子表达的影响。方法构建miR-146a过表达及抑制表达慢病毒载体并转染THP-1细胞,以正常培养的THP-1细胞为对照。用实时定量PCR法检测THP-1细胞AUF1 mRNA表达;蛋白质印迹法检测THP-1细胞AUF1蛋白的表达;酶联免疫吸附试验检测THP-1细胞培养基上清液白细胞介素-8(IL-8)及白细胞介素-35(IL-35)浓度。结果过表达miR-146a,可导致THP-1细胞AUF1 mRNA及蛋白表达下调(P<0.01,P<0.05);THP-1细胞上清液IL-8及IL-35浓度降低(P<0.01,P<0.05)。抑制miR-146a表达,导致THP-1细胞上清液IL-8及IL-35浓度明显增高(P<0.01,P<0.01)。结论 AUF1是miR-146a的靶基因。miR-146a可以调控THP-1细胞上清液IL-8、IL-35浓度。IL-8的改变引起相应IL-35的改变,一起参与炎性反应。

Objective To investigate the role of miR-146 a in regulating cytokine expression and the possible underlying mechanisms involved. Methods miR-146 a mimics or inhibitor were transfected into human acute monocytic leukemia cell line（ THP-1 cell line） by lentiviral vectors,and THP-1 cell in normal culture was used as control group. The expression levels of AUF1 in THP-1 cell lines were dectected by real-time PCR and Western blot. The levels of interleukin-8（ IL-8） and interleukin-35（ IL-35） in the culture supernatant were dectected by ELISA.Results miR-146 a mimics decreased,while miR-146 a inhibitor did not affect the expression of AUF1 in IL-1β-induced THP-1 cell lines. In addition,miR-146 a mimics decreased（ P〈 0. 01,P〈 0. 05）,while miR-146 a inhibitor increased（ P〈 0. 01,P〈 0. 01）,the expression of inflammatory cytokine IL-8 and anti-flammatory cytokine IL-35. Conclusions AUF1 is one of targets of miR-146 a. miR-146 a regulates the pro-inflammatory cytokine IL-8secretion,which is accompanied by anti-flammatory cytokine IL-35.