microRNA-155通过靶向抑制C/EBP-β和TAB2调控趋化因子的表达

microRNA-155 regulates chemokines expression by targeting C/EBP-β and TAB2

目的明确microRNA(miR)-155对趋化因子CCL5、CXCL9、CXCL10、CCL20和CCL22表达的调控作用及其靶基因。方法培养RAW264.7细胞,向细胞内转染寡聚核苷酸(包括miR-ctrl、miR-155 mimics和miR-155 inhibitor)或靶基因保护子(包括C/EBP-β-TP^(miR-155)、TAB2-TP^(miR-155)和SOCS1-TP^(miR-155)),并用1μg/ml LPS刺激细胞活化,用RT-PCR、ELISA和Western blotting方法检测趋化因子及miR-155靶基因表达情况。结果 (1)miR-155抑制LPS活化的RAW264.7细胞分泌CXCL9、CXCL10和CCL20(P<0.05),而促进上述细胞分泌CCL22(P<0.05);(2)C/EBP-β和TAB2分别是miR-155调控CXCL9和CXCL10表达的靶基因。结论 miR-155可通过调控趋化因子的表达参与调控免疫炎症反应。

This study was designed to demonstrate the role of microRNA（miR）-155 in regulating the expression of chemokines（including CCL5, CXCL9, CXCL10, CCL20, and CCL22）, and to clarify the target gene of miR-155 that involved in this process. RAW264.7 cells were cultured, and transfected with doligonucleotides（including miR-ctrl, miR-155 mimics, and miR-155 inhibitor） or target protectors（including C/EBP-β-TP^（miR-155）,TAB2-TP^（miR-155）, and SOCS1-TP^（miR-155））, then stimulated with 1 μg/ml LPS. The expression of these chemokines and the target genes of miR-155 were detected by RT-PCR, ELISA, and Western blotting. Data showed miR-155 inhibited the expression of CXCL9, CXCL10, and CCL20 in LPS activated RAW264.7 cells, and promoted the expression of CCL22 in those cells（P〈0.05）. C/EBP-β and TAB2 are target genes of miR-155 that involved in regulating the expression of CXCL9 and CXCL10, respectively. These results indicated that miR-155 regulates immune-inflammatory response by controlling chemokines expression.