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.
Highlights
Cataract is a common visual impairment in elderly people and the leading cause of blindness globally [1]
The expression of E2F transcription factor 3 (E2F3) messenger RNA (mRNA) and protein was remarkably downregulated in cataract lens tissues compared with the corresponding normal counterparts (Figure 1C and D)
SRA01/04 cells were transfected with anti-miR-NC, anti-miR-378a-5p, and anti-miR-630 to explore the effects of miR-378a-5p and miR-630 on cataract cell proliferation, apoptosis, and epithelial-mesenchymal transition (EMT)
Summary
Cataract is a common visual impairment in elderly people and the leading cause of blindness globally [1]. The projected number of cataract patients will climb to more than 30 million by 2020 according to census data provided by the USA [2]. The risk factors of cataract are complicated, such as smoking, hypertension, obesity, diabetes, drug usage, and age [3,4,5]. In the current development of medical therapeutic strategies, cataract surgery remains the most effective treatment due to the recovery of the pupillary reflex and optimization of light transmittance [6]. Poor medical care in developing countries impede favorable therapeutic outcomes. It is imperative to develop alternative therapies for cataract
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