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Alcohol induces TGF‐beta through suppression of miR‐1946a
Author(s) -
Sueblinvong Viranuj,
Mills Stephen T,
Fan Xian
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.778.15
Subject(s) - luciferase , microrna , untranslated region , transfection , messenger rna , three prime untranslated region , downregulation and upregulation , medicine , transforming growth factor , cancer research , microbiology and biotechnology , biology , cell culture , gene , biochemistry , genetics
Rationale We previously showed that chronic alcohol ingestion promotes fibroproliferative disrepair following bleomycin‐induced acute lung injury in a mouse model and that is associated with exaggerated constitutive and injury‐provoked expression of TGFβ. Importantly, the aberrant expression of TGFβ has been identified as a proximal event that mediates the ‘alcoholic lung’ phenotype. However, the mechanisms by which alcohol induces TGFβ1 has not been identified. Several microRNAs (miR) have been shown to directly target TGFβ. Using an in silico analysis, we identified that miR‐1946a is predicted to directly bind to the 3′ untranslated region (UTR) of the TGFβ mRNA and thereby inhibit its transcription. We hypothesize that alcohol‐mediated miR‐1946a leading to an induction of TGFβ expression. Methods Mouse primary lungfibroblasts (PLFs) were cultured ± alcohol (60 mM) for 24–48 hrs, miR‐1946aexpression was quantified. In parallel, PLFs were cultured with miR‐1946amimic, miR‐1946a inhibitor, or appropriate controls in the presence of absent of alcohol, then assessed for TGFβ gene and protein expression. Lastly, to further comfirm the miR‐1946abinding site on TGFβ 3′UTR, PLFs were transfected with vector containing TGFβ3′UTR (or control clone) tagged with luciferase reporter and miR‐1946a mimic(or negative mimic), then analyzed for luciferase activity. Results Alcohol suppressed PLFs expression of miR‐1946a. Treatment of PLFs with miR‐1946a mimic suppressed alcohol‐induced TGFβ expression while miR‐1946a inhibitor induced TGFβ expression. Lastly, miR‐1946a mimic suppressed TGFβ 3′UTR luciferase activity. Conclusion These data suggest that miR‐1946a modulates TGFβ expression likely through direct interaction with TGFβ 3′UTR. These findings identify a novel mechanism by which alcohol induces TGFβ in the lung which would help us design therapeutic or preventative tools for alcoholic patients with or at risk of acute lung injury. Support or Funding Information NIH K08 AA021404‐01 This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .