摘要
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.
作者
马志军
刘维军
李琳
樊世明
任明
柴秀英
马玉兰
MA Zhijun;LIU Weijun;LI Lin;FAN Shiming;REN Ming;CHAI Xiuying;MA Yulan(Department 2 of Surgical Oncology,Affiliated Hospital of Qinghai University,Xining 810001,China;Department of Cardiology,Affiliated Hospital of Qinghai University,Xining 810001,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2018年第11期940-945,共6页
Immunological Journal
基金
青海省卫生计生委项目(2017-wjzdx-52)
青海省科技厅项目(2017-ZJ-921Q)