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Published on: March 5, 2019
miR-140-5p suppresses retinoblastoma cell proliferation, migration, and invasion by targeting CEMIP and CADM3
Xiaolin Miao1, Zhen Wang2, Bingyu Chen2
1Outpatient Department, Eye Hospital of Wenzhou Medical University, China.
Abstract:
Retinoblastoma (RB) is a childhood intraocular tumor, affecting millions of patients worldwide. MicroRNA-140-5p (miR-140-5p) was demonstrated to be involved in the tumorigenesis of various human cancers; however, its role in RB remains undetermined. In this study, quantitative real-time PCR (qRT-PCR) and Western blot assays were used to determine the expression levels of miR-140-5p, cell migration-inducing protein (CEMIP), and cell adhesion molecule 3 (CADM3) in RB tissues and cell-lines. The proliferation ability was detected by cell-counting kit 8 (CCK-8), Edu staining, and colony formation assay. The cell cycle and migration and invasion abilities were measured by flow cytometry, wound-healing assay and Transwell assays, respectively. The correlation between miR-140-5p and CEMIP/CADM3 were then confirmed by immunofluorescence (IF) and dual-luciferase reporter assays. The results showed that miR-140-5p expression was significantly decreased; however, CEMIP and CADM3 expression was increased in RB tissues and cells. Overexpression of miR-140-5p inhibited proliferation, migration, and invasion of RB cells. We also found that miR-140-5p inhibited CEMIP and CADM3 expressions in RB cells. In addition, we demonstrated that miR-140-5p might negatively regulate the transcriptional activities of CEMIP and CADM3 by targeting their 3'-UTR. Therefore, we suggested that miR-140-5p could be a potential therapeutic target for the treatment of RB through CEMIP and CADM3.
Insights
MicroRNA-140-5p (miR-140-5p) is downregulated in retinoblastoma (RB), a childhood eye cancer. Restoring miR-140-5p inhibits RB cell growth and metastasis by targeting CEMIP and CADM3.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Retinoblastoma (RB) is a significant childhood intraocular tumor globally.
- The role of microRNA-140-5p (miR-140-5p) in RB tumorigenesis is currently unknown.
- Investigating miR-140-5p's function is crucial for understanding RB development.
Purpose of the Study:
- To determine the expression levels of miR-140-5p, CEMIP, and CADM3 in RB.
- To elucidate the functional role of miR-140-5p in RB cell proliferation, migration, and invasion.
- To explore the regulatory relationship between miR-140-5p and its potential targets, CEMIP and CADM3.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) and Western blot for gene/protein expression.
- Cell proliferation assays (CCK-8, Edu staining, colony formation).
- Cell cycle, migration, and invasion assays (flow cytometry, wound-healing, Transwell).
- Immunofluorescence (IF) and dual-luciferase reporter assays for target validation.
Main Results:
- miR-140-5p expression was significantly decreased in RB tissues and cell lines.
- CEMIP and CADM3 expression levels were inversely correlated with miR-140-5p.
- Overexpression of miR-140-5p suppressed RB cell proliferation, migration, and invasion.
- miR-140-5p was confirmed to target and inhibit CEMIP and CADM3 expression.
Conclusions:
- miR-140-5p acts as a tumor suppressor in retinoblastoma.
- The miR-140-5p/CEMIP/CADM3 axis plays a critical role in RB progression.
- miR-140-5p represents a potential therapeutic target for RB treatment.
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