Regulation of Apoptosis by miR-122 in Pterygium via Targeting Bcl-w

Invest Ophthalmol Vis Sci. 2016 Jul 1;57(8):3723-30. doi: 10.1167/iovs.16-19402.

Abstract

Purpose: To identify the differently expressed micro (mi) RNAs in pterygium compared with normal conjunctiva and investigate the potential role of miRNAs in the pathogenesis of pterygium.

Methods: With microRNA microarray and quantitative RT-PCR, we identified that microRNA-122 (miR-122) was significantly decreased in pterygium tissue. We detected the expression of Bcl-w, a predicted target of miR-122, in both pterygium and normal conjunctiva, as well as its correlation with the expression of miR-122. Pterygium epithelial cells were isolated and cultured, and transfected with miR-122 mimic or miR-122 inhibitor to change the miR-122 levels. The regulation of Bcl-w expression by miR-122 was examined with luciferase activity assay, quantitative (q) RT-PCR, and Western blot. The effect of the miR-122 on the apoptosis of cultured pterygium epithelial cells was investigated with TUNEL staining and caspase activity assay.

Results: We found the expression of Bcl-w, with an inverse correlation with the expression of miR-122, was significantly increased in pterygium, especially in the superficial layer of epithelium. In cultured pterygium epithelial cells, miR-122 could specifically combine with Bcl-w mRNA, and negatively regulated the expression of Bcl-w. Suppression of miR-122 could reduce apoptosis and caspase activity in pterygium epithelial cell treated with TNFα/cycloheximide (CHX), and this effect was abolished by inhibition of the expression of Bcl-w with specific siRNA.

Conclusions: Decreased expression of miR-122 in pterygium might result in abnormal cell apoptosis via its regulation of the expression of Bcl-w, and subsequently contribute to the development of pterygium.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Analysis of Variance
  • Apoptosis / physiology*
  • Apoptosis Regulatory Proteins / metabolism
  • Case-Control Studies
  • Caspases / metabolism
  • Cells, Cultured
  • Conjunctiva / metabolism
  • Cycloheximide / pharmacology
  • Down-Regulation / physiology
  • Epithelial Cells / metabolism
  • Humans
  • MicroRNAs / metabolism
  • MicroRNAs / physiology*
  • Protein Synthesis Inhibitors / pharmacology
  • Pterygium / etiology*
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • Apoptosis Regulatory Proteins
  • BCL2L2 protein, human
  • MIRN122 microRNA, human
  • MicroRNAs
  • Protein Synthesis Inhibitors
  • Tumor Necrosis Factor-alpha
  • Cycloheximide
  • Caspases