Photocoagulation of human retinal pigment epithelium in vitro: unravelling the effects on ARPE-19 by transcriptomics and proteomics

Acta Ophthalmol. 2015 Jun;93(4):348-54. doi: 10.1111/aos.12649. Epub 2015 Jan 21.

Abstract

Purpose: Despite the extensive use of retinal photocoagulation for ischaemia and vascular leakage in retinal vascular disease, the molecular mechanisms behind its clinical beneficial effects are still poorly understood. One important target of laser irradiation is the retinal pigment epithelium (RPE). In this study, we aimed at identifying the isolated effects of photocoagulation of RPE at both the mRNA and protein expression levels.

Methods: Human ARPE-19 cells were exposed to photocoagulation. Gene expression and protein expression were compared to untreated cells using microarray and liquid chromatography-mass spectrometry analysis. Genes and proteins queried by microarray and mass spectrometry were subjected to the Kyoto Encyclopedia of Genes and Genomes (KEGG) database pathway analyses.

Results: Laser irradiation resulted in an induction of the cytoprotective heat-shock protein subfamily Hsp70 as well as in a suppression of the vascular permeability factor carbonic anhydrase 9 (CA9). These expression patterns were evident at both the mRNA and protein levels. KEGG pathway analyses revealed genes and proteins involved in cellular turnover, repair and inflammation.

Conclusions: By characterizing the transcriptional and translational effects of laser coagulation on the RPE cells in culture, we have revealed responses, which might contribute to some of the beneficial effects obtained by photocoagulation for ischaemia and vascular leakage in retinal vascular disease.

Keywords: microarray; photocoagulation; proteomics; retinal pigment epithelium.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Line
  • Chromatography, High Pressure Liquid
  • Eye Proteins / genetics*
  • Eye Proteins / metabolism*
  • Gene Expression Profiling*
  • Humans
  • Laser Coagulation*
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Oligonucleotide Array Sequence Analysis
  • Proteomics*
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Retinal Pigment Epithelium / metabolism
  • Retinal Pigment Epithelium / surgery*
  • Tandem Mass Spectrometry
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transcriptome*

Substances

  • Eye Proteins
  • RNA, Messenger
  • Transcription Factors

Associated data

  • GENBANK/GSE41332