MicroRNA-30a Promotes B Cell Hyperactivity in Patients With Systemic Lupus Erythematosus by Direct Interaction With Lyn
MicroRNA-30a Promotes B Cell Hyperactivity in Patients With Systemic Lupus Erythematosus by Direct Interaction With Lyn
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DOI：
10.1002/art.37912
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发表时间：
2013-06-01
影响因子：
--
通讯作者：
Yang, Guang
中科院分区：
文献类型：
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作者：
Liu, Yu;Dong, Jie;Yang, Guang
目的探讨大多数系统性红斑狼疮（SLE）患者B细胞中Lyn水平显著降低的原因，并确定microRNA-30a （miR-30a）在SLE B细胞高活性中的作用。方法采用荧光素酶报告基因检测，鉴定miR-30a与Lyn 3-非翻译区（3-UTR）之间的相互作用。采用TaqMan定量聚合酶链式反应（qPCR）检测B细胞中miR-30a水平，采用实时荧光定量pcr检测Lyn信使RNA水平，采用Western blotting检测Lyn蛋白水平。采用酶联免疫吸附法测定IgG的含量。用3h -胸腺嘧啶掺入法测定B细胞的增殖。结果在B细胞系中，miR-30a可以特异性结合Lyn的3-UTR，而miR-30b、miR-30c、miR-30d和miR-30e则不能，过表达miR-30a可以抑制Lyn的水平。SLE患者B细胞中miR-30a水平明显高于健康供者。B细胞中miR-30a水平与Lyn水平呈负相关。发现过表达miR-30a可促进B细胞增殖和IgG抗体的产生。miR-30a的作用可以通过诱导Lyn在B细胞中过表达来消除。这些结果表明，miR-30a的表达升高是SLE患者B细胞中Lyn水平降低的原因，表明miR-30a在B细胞高活性中起重要作用。
Objective To investigate why the level of Lyn is significantly decreased in B cells from a majority of patients with systemic lupus erythematosus (SLE) and to determine the role of microRNA-30a (miR-30a) in SLE B cell hyperactivity. Methods Luciferase reporter gene assays were performed to identify the interaction between miR-30a and the 3-untranslated region (3-UTR) of Lyn. Levels of miR-30a in B cells were determined by TaqMan quantitative polymerase chain reaction (qPCR), Lyn messenger RNA levels were tested with real-time qPCR, and protein levels of Lyn were determined using Western blotting. The quantity of IgG was determined by enzyme-linked immunosorbent assay. The proliferation of B cells was measured using 3H-thymidine incorporation. Results In B cell lines, miR-30a, but not miR-30b, miR-30c, miR-30d, or miR-30e, could specifically bind the 3-UTR of Lyn, and overexpression of miR-30a inhibited the levels of Lyn. The level of miR-30a in B cells was significantly higher in SLE patients compared to healthy donors. The level of miR-30a was negatively associated with the level of Lyn in B cells. Overexpression of miR-30a was found to promote B cell proliferation and the production of IgG antibodies. The effect of miR-30a could be abrogated by inducing overexpression of Lyn in B cells. Conclusion These results reveal that elevated expression of miR-30a is responsible for the reduction in levels of Lyn in B cells from patients with SLE, suggesting that miR-30a plays an important role in B cell hyperactivity.