miR-708 and miR-335-3p Inhibit the Apoptosis of Retinal Ganglion Cells Through Suppressing Autophagy

J Mol Neurosci. 2021 Feb;71(2):284-292. doi: 10.1007/s12031-020-01648-y. Epub 2020 Jul 19.

Abstract

This study aimed to clarify the regulation role of miR-708 and miR-335-3p in retinal ganglion cell (RGC) autophagy and apoptosis in glaucoma. Chronic glaucoma mice were established by laser photocoagulation. RGCs were isolated and transfected with a series of plasmids and the cultured in 60 mmHg pressure. miR-335-3p, miR-708, and ATG3 mRNA expressions were detected by qRT-PCR. Protein levels of ATG3, autophagy-related protein LC3, and p62 were detected by Western blot. The apoptosis of RGCs was detected by flow cytometry. The regulation role of miR-335-3p/miR-708 in ATG3 was confirmed by the dual-luciferase reporter gene. The expressions of several miRNAs were measured in retinal tissues from chronic glaucoma mice and RGCs under pressure conditions, and results showed that both miR-335-3p and miR-708 were down-regulated. Besides, the inhibition of miR-708 and miR-335-3p induced the apoptosis of RGCs through promoting autophagy. Also, miR-708 and miR-335-3p could bind to ATG3 and targeted regulated ATG3. Furthermore, the interference with miR-708/miR-335-3p induced RGC apoptosis by up-regulating ATG3 to promote autophagy. In general, the down-regulation of miR-708 and miR-335-3p contributed to the apoptosis of RGCs through promoting autophagy in glaucoma.

Keywords: ATG3; Apoptosis; Autophagy; Glaucoma; Retinal ganglion cells; miR-335-3p; miR-708.

MeSH terms

  • Animals
  • Apoptosis / genetics*
  • Autophagy / genetics*
  • Autophagy-Related Proteins / biosynthesis
  • Autophagy-Related Proteins / genetics
  • Cells, Cultured
  • Down-Regulation
  • Glaucoma / genetics
  • Glaucoma / metabolism
  • Intraocular Pressure
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / genetics
  • MicroRNAs / physiology*
  • Microtubule-Associated Proteins / biosynthesis
  • Microtubule-Associated Proteins / genetics
  • Pressure
  • RNA / metabolism
  • RNA Interference
  • RNA, Messenger / genetics
  • RNA, Small Interfering / genetics
  • Retinal Ganglion Cells / cytology*
  • Ubiquitin-Conjugating Enzymes / biosynthesis
  • Ubiquitin-Conjugating Enzymes / genetics

Substances

  • Autophagy-Related Proteins
  • Map1lc3b protein, mouse
  • MicroRNAs
  • Microtubule-Associated Proteins
  • Mirn335 microRNA, mouse
  • RNA, Messenger
  • RNA, Small Interfering
  • RNA, recombinant
  • mirn708 microRNA, mouse
  • RNA
  • Ubiquitin-Conjugating Enzymes
  • Atg3 protein, mouse