Modulation of TLR2 protein expression by miR-105 in human oral keratinocytes.
Mammalian biological processes such as inflammation, involve regulation of hundreds of genes controlling onset and termination. MicroRNAs (miRNAs) can translationally repress target mRNAs and regulate innate immune responses. Our model system comprised primary human keratinocytes, which exhibited robust differences in inflammatory cytokine production (interleukin-6 and tumor necrosis factor-alpha) following specific Toll-like receptor 2 and 4 (TLR-2/TLR-4) agonist challenge. We challenged these primary cells with Porphyromonas gingivalis (a Gram-negative bacterium that triggers TLR-2 and TLR-4) and performed miRNA expression profiling. We identified miRNA (miR)-105 as a modulator of TLR-2 protein translation in human gingival keratinocytes. There was a strong inverse correlation between cells that had high cytokine responses following TLR-2 agonist challenge and miR-105 levels. Knock-in and knock-down of miR-105 confirmed this inverse relationship. In silico analysis predicted that miR-105 had complementarity for TLR-2 mRNA, and the luciferase reporter assay verified this. Further understanding of the role of miRNA in host responses may elucidate disease susceptibility and suggest new anti-inflammatory therapeutics.
Benakanakere
Manjunatha R
MR
Center for Oral Health and Systemic Disease, University of Louisville School of Dentistry, Louisville, Kentucky 40202, USA.
Li
Qiyan
Q
Eskan
Mehmet A
MA
Singh
Amar V
AV
Zhao
Jiawei
J
Galicia
Johnah C
JC
Stathopoulou
Panagiota
P
Knudsen
Thomas B
TB
Kinane
Denis F
DF
D015153:Blotting, Western; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D053762:Interleukin-12 Subunit p40 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D009841:Oligonucleotides / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D051195:Toll-Like Receptor 2 / Q000235:genetics* / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D053762:Interleukin-12 Subunit p40; D015850:Interleukin-6; C588223:MIRN105 microRNA, human; D035683:MicroRNAs; D009841:Oligonucleotides; D051195:Toll-Like Receptor 2; D014409:Tumor Necrosis Factor-alpha