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CircCUL2 suppresses retinoblastoma cells by regulating miR-214-5p/E2F2 Axis
Hui Zhang1, XinWen Qiu2, ZhiJie Song2
1Fuzhou Aier Eye Hospital, Fuzhou.
Abstract:
The aim of this study was to investigate the effect of circCUL2 on the proliferation, invasion and migration of retinoblastoma cells by regulating the miR-214-5p/E2F2 axis. qRT-PCR and western blot were performed to detect the expressions of circCUL2, miR-214-5p and E2F2 in tumor tissues and adjacent normal tissues from retinoblastoma patients, and in normal human retinal epithelial cells ARPE-19 and human retinoblastoma cells Y79 and SO-Rb50. qRT-PCR and western blot were performed for the detection of RNA levels of circCUL2 and miR-214-5p and the mRNA and protein levels of E2F2, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay for cell proliferation ability, Transwell assay for cell invasion ability, and scratch assay for cell migration ability. Luciferase dual reporter assay was used to detect the targeting relationship between circCUL2 and miR-214-5p, and between miR-214-5p and E2F2. CircCUL2 and E2F2 were lowly expressed, while miR-214-5p was highly expressed in retinoblastoma tumor tissues and cells. Transfection with pcDNA3.1-CUL2 or miR-214-5p inhibitor inhibited the proliferation, invasion and migration of Y79 and SO-Rb50 cells compared with the negative control; while transfection with sh-CUL2 or miR-214-5p mimics promoted the proliferation, invasion and migration of Y79 and SO-Rb50 cells. CircCUL2 negatively regulated miR-214-5p, while miR-214-5p negatively regulated E2F2. Overexpression of miR-214-5p or silencing of E2F2 in SO-Rb50 cells partially reversed the inhibitory effect of circCUL2 on the proliferation, invasion and migration of retinoblastoma cells. CircCUL2 inhibited the proliferation, invasion and migration of retinoblastoma cells by regulating the miR-214-5p/E2F2 axis.
Insights
Circular RNA CUL2 (circCUL2) inhibits retinoblastoma cell growth by regulating the miR-214-5p/E2F2 pathway. This study reveals circCUL2 as a potential therapeutic target for retinoblastoma, impacting cell proliferation, invasion, and migration.
Area of Science:
- Oncology
- Molecular Biology
- Ophthalmology
Background:
- Retinoblastoma is a common intraocular malignancy in children.
- Understanding the molecular mechanisms underlying retinoblastoma progression is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the role of circular RNA CUL2 (circCUL2) in retinoblastoma.
- To elucidate the regulatory axis of circCUL2, microRNA-214-5p (miR-214-5p), and E2F2 in retinoblastoma cells.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) and Western blot to assess gene and protein expression.
- Cell proliferation, invasion, and migration assays (MTT, Transwell, scratch assays).
- Luciferase dual-reporter assay to confirm targeting interactions.
Main Results:
- circCUL2 and E2F2 were downregulated, while miR-214-5p was upregulated in retinoblastoma tissues and cells.
- circCUL2 negatively regulated miR-214-5p, and miR-214-5p negatively regulated E2F2.
- circCUL2 suppressed retinoblastoma cell proliferation, invasion, and migration via the miR-214-5p/E2F2 axis.
Conclusions:
- circCUL2 acts as a tumor suppressor in retinoblastoma.
- The circCUL2/miR-214-5p/E2F2 pathway is a key regulator of retinoblastoma progression.
- circCUL2 holds potential as a therapeutic target for retinoblastoma treatment.
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