Long non-coding RNA PWRN2 regulates cytotoxicity in an in vitro model of age-related macular degeneration

Biochem Biophys Res Commun. 2021 Jan 8:535:39-46. doi: 10.1016/j.bbrc.2020.10.104. Epub 2020 Dec 16.

Abstract

Background: Age-related macular degeneration (AMD) may lead to irreversibly vision loss among aging populations. In this work, in an in vitro AMD cell model, we examined the expression and function of long non-coding RNA, Prader-Willi Region Non-Protein Coding RNA 2 (PWRN2) in injured human retinal pigment epithelial cells.

Method: ARPE-19 cell line was maintained in vitro and treated with multi-module stressful conditions, including hydrogen peroxide (H2O2) tert-butylhydroperoxide (t-BuOOH) and ultraviolet B (UVB). Multi-module-stressor-induced cell death was monitored by a viability assay, and PWRN2 expression by qRT-PCR. PWRN2 was either downregulated or upregulated in ARPE-19 cells. The effects of PWRN2 downregulation or upregulation on t-BuOOH-induced cell death, cellular apoptosis and mitochondrial injuries were then quantitatively evaluated.

Results: Multi-module stressful conditions induced cell death and PWRN2 upregulation in ARPE-19 cells in vitro. We created ARPE-19 subpopulations with either downregulated or upregulated PWRN2 expressions. Quantitative assays demonstrated that, PWRN2 downregulation effectively alleviated t-BuOOH-induced cell death, apoptosis and various-type of mitochondrial injuries. On the other hand, PWRN2 upregulation worsened t-BuOOH-induced cellular damages in ARPE-19 cells.

Conclusion: We demonstrated that downregulating PWRN2 protected multi-module-stressor-induced cell death, apoptosis and mitochondrial injuries in human retinal pigment epithelial cells, suggesting PWRN2 may be an active factor in human AMD.

Keywords: AMD; Apoptosis; Cell death; PWRN2; Retina; lncRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Death
  • Cell Line
  • Cell Survival / genetics
  • Down-Regulation / genetics
  • Epithelial Cells / metabolism
  • Epithelial Cells / pathology
  • Humans
  • Macular Degeneration / genetics*
  • Macular Degeneration / pathology
  • Mitochondria / metabolism
  • Models, Biological*
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism*
  • Retinal Pigment Epithelium / pathology
  • Up-Regulation / genetics
  • tert-Butylhydroperoxide

Substances

  • RNA, Long Noncoding
  • tert-Butylhydroperoxide