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Inhibitory effect of miR-145 on RPE cell proliferation
Ken Zhao1, Zhen Chen2, Xu-Dong Lv3
1Department of Ophthalmology, The Second Affiliated Hospital of Hubei Polytechnic College; The People's Hospital of Daye City Daye 435100, Hubei Province, P. R. China.
Objective:
This study aims to explore the impact of micro RNA miR-145 on retinal pigment epithelial cell proliferation and apoptosis.
Methods:
A stable culture and passage system of hPNE cells was first established, and its migration ability was determined. Then, miR-145 lentiviral vectors were constructed to transfect hPRE cells. Thereafter, hRPE cell proliferation was detected by MTT assay after they were transfected by lentivirus, cell cycle was analyzed by flow cytometry, and apoptosis was detected by Annexin V/PI double staining immunofluorescence.
Results:
Cultured hPRE cells had good migrating and metastatic ability, in which subsequent lentivirus infection experiments can be carried out. After transfection by miR-145 lentiviral vectors, hPRE cell proliferation slowed down and RPE cells in the G1 phase was inhibited; thus, apoptosis rate increased.
Conclusion:
MiR-145 can slow down retinal pigment epithelial cell proliferation and increase their apoptosis rate. This has a certain therapeutic potential for diseases caused by RPE cell proliferation such as PVR.
Insights
MicroRNA miR-145 inhibits retinal pigment epithelial cell proliferation and promotes apoptosis. This finding suggests miR-145 has therapeutic potential for proliferative vitreoretinopathy (PVR).
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Retinal pigment epithelial (RPE) cells play a crucial role in retinal health.
- Dysregulated RPE cell proliferation is implicated in blinding diseases like proliferative vitreoretinopathy (PVR).
- MicroRNAs (miRNAs) are key regulators of cellular processes, including proliferation and apoptosis.
Purpose of the Study:
- To investigate the effect of microRNA miR-145 on the proliferation and apoptosis of human retinal pigment epithelial (hRPE) cells.
- To assess the potential of miR-145 as a therapeutic agent for RPE-related proliferative diseases.
Main Methods:
- Established a stable culture system for hRPE cells and assessed their migratory capacity.
- Constructed miR-145 lentiviral vectors for transfection into hRPE cells.
- Utilized MTT assays, cell cycle analysis (flow cytometry), and Annexin V/PI staining to evaluate proliferation and apoptosis.
Main Results:
- Transfection with miR-145 lentiviral vectors significantly reduced hRPE cell proliferation.
- miR-145 induced cell cycle arrest in the G1 phase.
- An increased rate of apoptosis was observed in hRPE cells following miR-145 transfection.
Conclusions:
- MiR-145 effectively inhibits retinal pigment epithelial cell proliferation and enhances apoptosis.
- These findings indicate that miR-145 holds therapeutic promise for conditions characterized by excessive RPE cell proliferation, such as PVR.
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