April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
MicroRNA-29b Regulates TGF-β1-Mediated Epithelial-Mesenchymal Transition of Retinal Pigment Epithelial Cells by Targeting AKT2
Author Affiliations & Notes
  • Fang Wang
    Department of Ophthalmology, Shanghai Tenth People, Shanghai, China
    Eye Institute, Tongji University, Shanghai, China
  • Min Li
    Department of Ophthalmology, Shanghai Tenth People, Shanghai, China
  • Hui Li
    Department of Ophthalmology, Shanghai Tenth People, Shanghai, China
  • Chen Sun
    Department of Ophthalmology, Shanghai Tenth People, Shanghai, China
    Eye Institute, Tongji University, Shanghai, China
  • Hao Wang
    Department of Ophthalmology, Shanghai Tenth People, Shanghai, China
  • Jun Yao
    Eye Institute, Tongji University, Shanghai, China
  • Footnotes
    Commercial Relationships Fang Wang, None; Min Li, None; Hui Li, None; Chen Sun, None; Hao Wang, None; Jun Yao, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 4994. doi:
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      Fang Wang, Min Li, Hui Li, Chen Sun, Hao Wang, Jun Yao, ; MicroRNA-29b Regulates TGF-β1-Mediated Epithelial-Mesenchymal Transition of Retinal Pigment Epithelial Cells by Targeting AKT2. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4994.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

The role of microRNA (miRNA) in proliferative vitreoretinopathy (PVR) progression has not been studied extensively, especially in retinal pigment epithelial-mesenchymal transition (EMT) which is the main reason for formation of PVR. The aim of this study was to investigate and validate unidentified miRNAs that regulate EMT and to reveal the function in PVR clinical samples.

 
Methods
 

MiRNA expression profile in ARPE-19 cells EMT model was performed by microRNA microarray. Immunofluorescence staining, Q-PCR, western blot were conducted to determine the level of three epithelial and mesenchymal markers zona occludin-1 (ZO-1), E-cadherin and α-smooth muscle actin (α-SMA) in ARPE-19 cells and PVR membranes. The migration ability of treated ARPE-19 cells was accessed by Transwell migration assay. The target of miR-29b was predicted by computational approach and identified by Dual luciferase activity assay.

 
Results
 

MiRNA microarray detected that miR-3138, miR-22*, and miR-455-3p were upregulated, while miR-550a, miR-135b and miR-29b were significantly downregulated in TGF-β1 mediated EMT of ARPE-19 cells. Among the five changed miRNAs, miR-29b showed the most significant downregulation. Enhanced expression of miR-29b could reverse TGF-β1 induced EMT through targeting Akt2(Figure1 and 2). Akt2 downregulation could inhibit TGF-β1 induced EMT. Furthermore, inhibition of miR-29b in ARPE-19 cells directly triggered EMT process, which characterized by the phenotypic transition and the upregulation of α-SMA and downregulation of E-cadherin and ZO-1 with increased cell migration. Akt2-shRNA also inhibited miR-29 inhibitor induced EMT process. In epiretinal membrane and vitreous of PVR patients, miR-29b expression was downregulated and positively correlated with E-cadherin and ZO-1 mRNA expression, while an inverse correlation was observed between miR-29b and α-SMA or Akt2 mRNA expression.

 
Conclusions
 

Our findings suggest that miR-29b plays an important role in TGF-β1-mediated EMT in ARPE-19 cells and PVR pathological process by targeting Akt2.

 
 
Figure 1 miR-29b inhibited TGF-β1 induced EMT.
 
Figure 1 miR-29b inhibited TGF-β1 induced EMT.
 
 
Figure 2 MiR-29b targeted Akt2 in ARPE-19 cells.
 
Figure 2 MiR-29b targeted Akt2 in ARPE-19 cells.
 
Keywords: 655 proliferative vitreoretinopathy • 701 retinal pigment epithelium  
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