title: [miR-338-5p modulates B cell biological functions by targeting NF-κB1].
abstract: Objective To explore the effects of miR-338-5p on the nuclear factor κB1 (NF-κB1) expression and the IgG-producing ability of B cells. Methods Dual-luciferase reporter assay was used to test the target gene of miR-338-5p. The purified CD20+B cells were transfected with miR-338-5p agomiR, miR-338-5p antagomiR, NF-κB1 siRNA (siNF-κB1) and their corresponding negative control reagents, and then cultured with anti-IgM antibody and/or recombinant human B cell activating factor (rhBAFF). Real-time RCR and Western boltting were applied to determine the mRNA and protein levels of NF-κB1. IgG level in the supernatant was detected by ELISA. Results Compared with the control group, the hRluc/hLuc relative luciferase activity was significantly elevated in miR-338-5p mimic and NF-κB1-3'-UTR reporter co-transfected group. In the co-culture system with anti-IgM antibody and rhBAFF, the NF-κB1 mRNA, p105, p50 and IgG levels in B cells transfected with miR-338-5p agomiR were significantly increased, while the NF-κB1 mRNA and IgG levels in B cells transfected with miR-338-5p antagomiR were significantly decreased. The effect of siNF-κB1 on B cells was opposite to that of miR-338-5p agomiR. Correlation analysis suggested that NF-κB1 mRNA level was significantly positively correlated with IgG concentration. Conclusion miR-338-5p regulates the biological functions of B cells by positively regulating NF-κB1 expression and indirectly regulating BAFF signal.
authors: Department of Urinary Surgery, Third Affiliated Hospital of Soochow University, Changzhou 213003, China.
authors: Xuyi
authors: X
authors: Ma
authors: Department of Urinary Surgery, Third Affiliated Hospital of Soochow University, Changzhou 213003, China.
authors: Xiaozhou
authors: X
authors: He
authors: Department of Urinary Surgery, Third Affiliated Hospital of Soochow University, Changzhou 213003, China.
authors: Panpan
authors: P
authors: Dong
authors: Department of Urinary Surgery, Third Affiliated Hospital of Soochow University, Changzhou 213003, China.
authors: Dong
authors: D
authors: Xue
authors: Department of Urinary Surgery, Third Affiliated Hospital of Soochow University, Changzhou 213003, China.
authors: Dan
authors: D
authors: Song
authors: Department of Urinary Surgery, Third Affiliated Hospital of Soochow University, Changzhou 213003, China. *Corresponding author, E-mail: xuhaiyan09@163.com.
authors: Haiyan
authors: H
authors: Xu
authors: Institute of Medical Biology, Soochow University, Suzhou 215007, China.
authors: Xueguang
authors: X
authors: Zhang
mesh_terms: D053264:B-Cell Activating Factor / Q000235:genetics / Q000378:metabolism; D001402:B-Lymphocytes / Q000378:metabolism*; D015153:Blotting, Western; D057809:HEK293 Cells; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection
chemical_list: D053264:B-Cell Activating Factor; D007074:Immunoglobulin G; C532050:MIRN338 microRNA, human; D035683:MicroRNAs; D052002:NF-kappa B p50 Subunit; C489548:NFKB1 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering