基础医学与临床 ›› 2016, Vol. 36 ›› Issue (12): 1675-1680.

• 研究论文 • 上一篇    下一篇

miR-146a对THP-1细胞靶基因AUF1调控及细胞因子表达的影响

马国添1,曾志羽1,夏炳辉2,林士云2,柯红红3,梁艺王莹2   

  1. 1. 广西医科大学第一附属医院
    2. 广西医科大学 第一附属医院
    3. 广西医科大学第一附属医院心内科
  • 收稿日期:2016-02-14 修回日期:2016-05-24 出版日期:2016-12-05 发布日期:2016-11-29
  • 通讯作者: 马国添 E-mail:drguotianma@163.com
  • 基金资助:
    广西医疗卫生适宜技术研究与开发课题;广西医科大学青年科学基金

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

  • Received:2016-02-14 Revised:2016-05-24 Online:2016-12-05 Published:2016-11-29

摘要: 目的 探讨miR-146a对人急性单核白血病细胞株(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。

关键词: 人急性单核白血病细胞株, 微小核糖核酸-146a, AU碱基富集区结合降解因子1, 细胞因子

Abstract: Objective To investigate the role of miR-146a in regulating cytokine expression and the possible underlying mechanisms involved. Methods miR-146a 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 were 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-146a mimics decreased, while miR-146a inhibitor did not affect the expression of AUF1 in IL-1β-induced THP-1 cell lines. In addition, miR-146a mimics decreased(P<0.01; P<0.05), while miR-146a 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-146a. miR-146a regulates the pro-inflammatory cytokine IL-8 secretion, which is accompanied by anti-flammatory cytokine IL-35.

Key words: THP-1 cell lines, MicroRNA-146a, AUF1, Cytokine

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