MicroRNA 144 negatively regulates Toll-like receptor 2 expression in rat macrophages
WANG Xuan
LAN Xi
LIU Li
YI Jing
LI Jing
LI Yue
WANG Meichen
LI Jiaxi
SONG Liumei
LI Dongmin
Abstract：Objective To investigate the relationship between miR-144 and Toll-like receptor 2 (TLR2). Methods RT-qPCR was used to determine the expression of TLR2 and its downstream inflammatory cytokine TNF-α in rat macrophage cell line NR8383 transfected by a mimic miR-144 or miR-144 inhibitor. The fragments of 3'UTR region of rat TLR2 mRNA including wild or mutant miR-144 binding site obtained by PCR using rat liver cDNA were ligated to pmirGLO report gene vector digested with SacI and XbaI to construct the recombinant vectors of pmir-TLR2-3'UTR and pmir-mutant-TLR2-3'UTR. The miR-144 targeting TLR2 was further determined by dual luciferase reporter assay and miR-144 mimics. Results TLR2 and TNF-αin NR8383 cells were decreased after transfection with 100 nmol/L mimic miR-144 for 24 h and increased after transfection with 100 nmol/L miR-144 inhibitor. PCR and double-enzyme digestion with SacI and XbaI confirmed successful insertion of the target fragments. Dual luciferase reporter assay suggested the binding of miR-144 to the 3'UTR of rat TLR2 mRNA. Conclusion miR-144 negatively regulates the expression of TLR2 and its down-stream cytokine TNF-αby targeting TLR2 in NR8383 cells.
Keywords：miR-144Toll-like receptor 2NR8383 cellsmacrophagePmirGLO report gene carrier
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 319-325 )