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Effect of NF-κB p65 antisense oligodeoxynucleotide on transdifferentiation of normal human lens epithelial cells

Chao Liu1, Xiao-Li Wu2, Xin-Yi Wu3

  • 1Department of Ophthalmology, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China; Department of Ophthalmology, Central Hospital of Qingdao, Qingdao 266042, Shandong Province, China.

International Journal of Ophthalmology
|March 8, 2016
PubMed
Abstract

Insights

Nuclear factor kappa-B p65 antisense oligodeoxynucleotide (NF-κB p65 ASODN) inhibited transforming growth factor-β2 (TGF-β2)-induced transdifferentiation in human lens epithelial cells. This suggests NF-κB p65 ASODN may be a therapeutic target for posterior capsular opacification.

Area of Science:

  • Ophthalmology
  • Cell Biology
  • Molecular Biology

Background:

  • Posterior capsular opacification (PCO) is a common complication after cataract surgery.
  • Transforming growth factor-β2 (TGF-β2) is implicated in the transdifferentiation of lens epithelial cells, a key process in PCO development.
  • Nuclear factor kappa-B p65 (NF-κB p65) signaling pathway activation is observed during TGF-β2-induced transdifferentiation.

Purpose of the Study:

  • To investigate the inhibitory effect of NF-κB p65 antisense oligodeoxynucleotide (ASODN) on TGF-β2-induced transdifferentiation of human lens epithelial cells (HLE B-3) in vitro.
  • To evaluate NF-κB p65 ASODN as a potential therapeutic target for PCO.

Main Methods:

  • Human lens epithelial cells (HLE B-3) were treated with TGF-β2 to induce transdifferentiation.
  • Cells were transfected with NF-κB p65 ASODN or a missense control oligodeoxynucleotide (MSODN).
  • Expression levels of NF-κB p65 mRNA and α-smooth muscle actin (α-SMA) protein were quantified using RT-PCR and ELISA, respectively.

Main Results:

  • TGF-β2 significantly increased the expression of NF-κB p65 mRNA and α-SMA protein in a time-dependent manner.
  • NF-κB p65 ASODN treatment significantly reduced the expression of both NF-κB p65 mRNA and α-SMA protein induced by TGF-β2.
  • NF-κB p65 MSODN and the transfer agent HiPerFect did not show significant inhibitory effects.

Conclusions:

  • NF-κB p65 ASODN effectively antagonizes TGF-β2-induced transdifferentiation of human lens epithelial cells in vitro.
  • The findings support NF-κB p65 ASODN as a promising therapeutic target for preventing or treating posterior capsular opacification.
Keywords:
antisense oligodeoxynucleotidelens epithelial cellsnuclear factor kappa-B p65transforming growth factor-β2α-smooth muscle actin

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