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Larry Estlack, Ginger Pocock, Brent Lavey, Adam Schenk, Jeffrey Wigle; Mir-146a Promotes Cell Proliferation And Survival By Targeting Nf-kb In A Human Retinal Pigment Epithelial Cell Line. Invest. Ophthalmol. Vis. Sci. 2012;53(14):5138.
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© ARVO (1962-2015); The Authors (2016-present)
To study the effect of low power laser inactivation (LPLI) on the expression of miR-146a in RPE cells in association with cell proliferation and survival by targeting NF-kB.
A human retinal pigment epithelial cell line (hTERT-RPE1) was exposed to LPLI (670 nm, 2.88 J/cm2). The cell line was then characterized to determine the relative abundance of protein and mRNA levels of NF-kB and the up-regulation (expression) of miR-146a. Cell proliferation and survival assays were used to determine the effects of miR-146a targeted NF-kB.
Following LPLI (670 nm, 2.88 J/cm2) expression kinetics for NFkB protein and mRNA levels were increased by 7 fold at 24 hours post exposure and 6 fold at 6 hours post exposure, respectively. Up-regulation of miR-146a was 10 fold at 2 hours post exposure. Compared to unexposed control cells the exposed cells had an increase of 25% in cell proliferation and survival 48 hours post exposure. Antagomir-mediated inhibition of miR-146a contributed to a decrease in expression of NFkB protein (24 hours post exposure) and mRNA (6 hours post exposure) by about 35% and 42%, respectively and the increase in cell proliferation and survival 48 hours post exposure was no longer visible.
Up-regulation of miR-146a induced by LPLI (670 nm, 2.88 J/cm2) targets the attenuation of NF-kB activity promoting cell proliferation and survival. This phenomenon represents a possible negative feedback loop affecting post transcriptional proteins. Therefore it may be possible to regulate RPE cell growth utilizing LPLI by affecting transcription controlled pathways.
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