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Published on: December 11, 2020
Hyperosmotic stress induces cell cycle arrest in retinal pigmented epithelial cells
T Arsenijevic1, A Vujovic, F Libert
1Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.
Abstract:
Osmotic changes occur in many tissues and profoundly influence cell function. Herein, we investigated the effect of hyperosmotic stress on retinal pigmented epithelial (RPE) cells using a microarray approach. Upon 4-h exposure to 100 mM NaCl or 200 mM sucrose, 79 genes were downregulated and 72 upregulated. Three gene ontology categories were significantly modulated: cell proliferation, transcription from RNA polymerase II promoter and response to abiotic stimulus. Fluorescent-activated cell sorting analysis further demonstrated that owing to hyperosmotic stimulation for 24 h, cell count and cell proliferation, as well as the percentage of cells in G0/G1 and S phases were significantly decreased, whereas the percentage of cells in G2/M phases increased, and apoptosis and necrosis remained unaffected. Accordingly, hyperosmotic conditions induced a decrease of cyclin B1 and D1 expression, and an activation of the p38 mitogen-activated protein kinase. In conclusion, our results demonstrate that hypertonic conditions profoundly affect RPE cell gene transcription regulating cell proliferation by downregulation cyclin D1 and cyclin B1 protein expression.
Insights
Hyperosmotic stress impacts retinal pigmented epithelial cells by altering gene expression and cell cycle progression. This stress reduces cell proliferation and affects key proteins involved in cell division.
Area of Science:
- Cell Biology
- Molecular Biology
- Ophthalmology
Background:
- Osmotic changes are critical in regulating tissue and cell function.
- Retinal Pigmented Epithelial (RPE) cells play a vital role in retinal health.
- Understanding RPE cell response to osmotic stress is crucial for ocular health research.
Purpose of the Study:
- To investigate the effects of hyperosmotic stress on RPE cells.
- To identify gene expression changes in RPE cells under osmotic stress.
- To elucidate the impact of hyperosmotic stress on RPE cell cycle and apoptosis.
Main Methods:
- Microarray analysis to assess gene expression changes.
- Exposure of RPE cells to hyperosmotic conditions (NaCl and sucrose).
- Fluorescent-activated cell sorting (FACS) for cell cycle and apoptosis analysis.
Main Results:
- Hyperosmotic stress significantly altered the expression of 151 genes (79 downregulated, 72 upregulated).
- Cell proliferation, cell count, and G0/G1 and S phases decreased, while G2/M phase increased.
- Apoptosis and necrosis remained unaffected; cyclin B1 and D1 expression decreased, and p38 MAPK was activated.
Conclusions:
- Hypertonic conditions profoundly affect RPE cell gene transcription.
- Cell proliferation is regulated by the downregulation of cyclin D1 and cyclin B1 protein expression.
- Hyperosmotic stress impacts RPE cell cycle progression without inducing cell death.
