Skip Navigation LinksHome > October 2013 - Volume 19 - Issue 11 > miR-106b Fine Tunes ATG16L1 Expression and Autophagic Activi...
Inflammatory Bowel Diseases:
doi: 10.1097/MIB.0b013e31829e71cf
Original Basic Science Articles

miR-106b Fine Tunes ATG16L1 Expression and Autophagic Activity in Intestinal Epithelial HCT116 Cells

Zhai, Zili PhD; Wu, Feng MD, PhD; Chuang, Alice Y. MD; Kwon, John H. MD, PhD

Supplemental Author Material
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Abstract

Background:

The microRNAs (miRNAs) regulate gene expression at the posttranscriptional level. ATG16L1, an essential component for autophagy and a risk gene for Crohn’s disease, contains two binding sites in the 3’UTR for miR-17 family, including miRs-20a, -93, -106a, and -106b. The purpose of this study was to assess the effects of these miRNAs on ATG16L1 expression and autophagic activity in HCT116 cells.

Methods:

The functional binding sites in the ATG16L1 3’UTR were evaluated by transfection of pMIR-GLO vectors bearing the wild type or mutant 3’UTR into cells for luciferase reporter assay. The miRNA regulation of ATG16L1 expression was determined by quantitative real-time polymerase chain reaction and Western blot. The miRNA regulation of autophagic activity was evaluated by examining LC3II formation using Western blot and confocal imaging.

Results:

Both miR-106a and miR-106b mimics inhibited starvation-induced autophagy. The miR-106b mimic reduced ATG16L1 protein expression. Luciferase reporter assays showed that mutating the binding sequence at the positions 1036 to 1042 abrogated miR-106b regulation of ATG16L1 3’UTR luciferase activity. In addition, miR-106a and miR-106b overexpression inhibited the expression of several other autophagy genes, including ATG12.

Conclusions:

miR-106b targets ATG16L1 and modulates autophagy, partially through the binding site at the 3’ end of ATG16L1 3’UTR. miR-106a regulates autophagy, possibly irrelevant to ATG16L1 regulation. Both miR-106a and miR-106b regulate multiple autophagy genes so that they may play an integral role in fine-tuning autophagy.

Copyright © 2013 Crohn's & Colitis Foundation of America, Inc.

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