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Circ_0000099/miR-223-3p/CTGF 通过调控 TGF-β2 刺激的人晶状体上皮细胞中的 EMT 过程来调节细胞的生长和转移。

Circ_0000099/miR-223-3p/CTGF Regulates the Growth, Metastasis, and EMT Processes in TGF-β2-Stimulated Human Lens Epithelial Cells.

机构信息

The Affiliated Nanhua Hospital, Department of Ophthalmology, Hengyang Medical School, University of South China, Hengyang, China.

出版信息

Curr Eye Res. 2024 Oct;49(10):1042-1053. doi: 10.1080/02713683.2024.2357600. Epub 2024 Jun 28.

DOI:10.1080/02713683.2024.2357600
PMID:38940233
Abstract

PURPOSE

Posterior capsule opacification (PCO) is the major complication of visual impairment after cataract surgery. Circular RNAs (circRNAs) are involved in the development of many diseases. The purpose of this study was to explore the role and molecular mechanism of circ_0000099 in PCO.

METHODS

SRA01/04 cells were treated with TGF-β2 to establish a PCO cell model. The expression of circ_0000099, miR-223-3p, and connective tissue growth factor (CTGF) mRNA was determined by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot assay was used to analyze the protein expression. Cell proliferation, migration, and invasion were analyzed by (4-5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), 5-ethynyl-2 '-Deoxyuridine (EdU), transwell, and wound healing tests. The circ_0000099/miR-223-3p/CTGF relationship was verified by dual luciferase reporter gene and RNA binding protein immunoprecipitation (RIP) assays.

RESULTS

TGF-β2 treatment promoted SRA01/04 cell proliferation invasion, migration, and EMT. Circ_0000099 expression was increased in POC patients and TGF-β2-treated SRA01/04 cells.Knockdown of circ_0000099 suppressed TGF-β2-induced proliferation, invasion, migration, and EMT in SRA01/04 cells. miR-223-3p was identified as the target of circ_0000099, and miR-223-3p inhibitor might partly abolish the repression of circ_0000099 silencing on TGF-β2-triggered SRA01/04 cell disorders. MiR-223-3p directly targeted CTGF. Knockdown of CTGF suppressed TGF-β2-induced SRA01/04 cell injury. Circ_0000099 can regulate CTGF expression by targeting miR-223-3p.

CONCLUSIONS

Circ_0000099 silencing might relieve TGF-2-induced SRA01/04 cell injury by the miR-223-3p/CTGF axis, providing new avenues for the prevention and treatment of PCO.

摘要

目的

后囊混浊(PCO)是白内障手术后视力损害的主要并发症。环状 RNA(circRNA)参与许多疾病的发展。本研究旨在探讨 circ_0000099 在 PCO 中的作用和分子机制。

方法

用 TGF-β2 处理 SRA01/04 细胞建立 PCO 细胞模型。实时定量聚合酶链反应(qRT-PCR)测定 circ_0000099、miR-223-3p 和结缔组织生长因子(CTGF)mRNA 的表达。Western blot 分析蛋白表达。四甲基偶氮唑蓝(MTT)、5-乙炔基-2 '-脱氧尿苷(EdU)、transwell 和划痕愈合试验分析细胞增殖、迁移和侵袭。通过双荧光素酶报告基因和 RNA 结合蛋白免疫沉淀(RIP)试验验证 circ_0000099/miR-223-3p/CTGF 关系。

结果

TGF-β2 处理促进 SRA01/04 细胞增殖、侵袭、迁移和 EMT。PCO 患者和 TGF-β2 处理的 SRA01/04 细胞中 circ_0000099 表达增加。circ_0000099 敲低抑制 SRA01/04 细胞中 TGF-β2 诱导的增殖、侵袭、迁移和 EMT。miR-223-3p 被鉴定为 circ_0000099 的靶标,miR-223-3p 抑制剂可能部分消除 circ_0000099 沉默对 TGF-β2 触发的 SRA01/04 细胞紊乱的抑制作用。miR-223-3p 直接靶向 CTGF。CTGF 敲低抑制 SRA01/04 细胞损伤。circ_0000099 通过靶向 miR-223-3p 调节 CTGF 表达。

结论

circ_0000099 沉默可能通过 miR-223-3p/CTGF 轴缓解 TGF-2 诱导的 SRA01/04 细胞损伤,为 PCO 的预防和治疗提供新途径。

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