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miR-378a-5p and miR-630 induce lens epithelial cell apoptosis in cataract via suppression of E2F3
Weiwei Gao1, Xiaoqing Zhou2, Ruihua Lin1
1Department of Ophthalmology, People's Hospital of Zhaoyuan City, Zhaoyuan, Shandong, China.
Abstract:
Cataract, an eye disease that threatens the health of millions of people, brings about severe economic burden for patients and society. MicroRNA (miR)-378a-5p and miR-630 were recognized as essential regulators in multiple cancers. However, the exact functions of miR-378a-5p and miR-630 in cataract are still unclear. The expression of miR-378a-5p, miR-630, and E2F transcription factor 3 (E2F3) in tissues and cells was measured by quantitative real-time polymerase chain reaction. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay was used to evaluate cell viability. Flow cytometry was conducted to analyze cell apoptosis. The interaction between E2F3 and miR-378a-5p or miR-630 was confirmed by dual-luciferase reporter assay. The expression of proteins E2F3, B cell lymphoma (Bcl-2), Bcl-2 associated X (Bax), and cleaved caspase 3 was detected by western blot assay. The expression of miR-378a-5p and miR-630 was up-regulated whereas E2F3 was down-regulated in human cataract lens tissues compared with normal lens tissues. Depletion of miR-378a-5p or miR-630 enhanced proliferation and reduced apoptosis of human lens epithelial cells. Interestingly, up-regulation of E2F3 exhibited the same trend. Next, dual-luciferase reporter assay validated the interaction between E2F3 and miR-378a-5p or miR-630. The rescue experiments further revealed that E2F3 knockdown could recover miR-378a-5p, and miR-630 inhibitor induced promotion of cell proliferation and inhibition of apoptosis in cataract. miR-378a-5p and miR-630 repressed proliferation and induced apoptosis of lens epithelial cells by targeting E2F3 in cataract, representing a prospective alternative therapy for cataract.
Insights
MicroRNA-378a-5p and microRNA-630 target E2F3 to inhibit lens cell proliferation and promote apoptosis in cataracts. These microRNAs offer potential new therapies for this common eye disease.
Area of Science:
- Ophthalmology
- Molecular Biology
- Genetics
Background:
- Cataract is a leading cause of vision loss globally, imposing significant economic and social burdens.
- MicroRNAs (miRNAs), specifically miR-378a-5p and miR-630, are known regulators in various cancers, but their role in cataract remains unexplored.
Purpose of the Study:
- To investigate the functional roles of miR-378a-5p and miR-630 in the pathogenesis of cataract.
- To elucidate the molecular mechanism by which these miRNAs regulate human lens epithelial cells.
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) to measure miRNA and E2F3 expression.
- Cell viability (MTT assay) and apoptosis (flow cytometry) assays.
- Dual-luciferase reporter assay to confirm miRNA-target interactions.
- Western blot to detect key protein expression (E2F3, Bcl-2, Bax, cleaved caspase 3).
Main Results:
- miR-378a-5p and miR-630 were upregulated, while E2F3 was downregulated in human cataract tissues.
- Depletion of miR-378a-5p or miR-630 increased lens epithelial cell proliferation and decreased apoptosis.
- E2F3 was validated as a direct target of both miR-378a-5p and miR-630.
- Restoration of E2F3 expression reversed the effects of miRNA depletion, confirming the regulatory pathway.
Conclusions:
- miR-378a-5p and miR-630 suppress lens epithelial cell proliferation and induce apoptosis by targeting E2F3 in cataract.
- These findings highlight miR-378a-5p and miR-630 as potential therapeutic targets for cataract treatment.
