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

IF 1.7 4区 医学 Q3 OPHTHALMOLOGY Current Eye Research Pub Date : 2024-10-01 Epub Date: 2024-06-28 DOI:10.1080/02713683.2024.2357600
Hong Tang, Shu Shu, Shiqin Hu, Le Chen
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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.

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Circ_0000099/miR-223-3p/CTGF调控TGF-β2刺激的人晶状体上皮细胞的生长、转移和EMT过程
目的:后囊不透明(PCO)是白内障手术后视力受损的主要并发症。环状 RNA(circRNA)参与了许多疾病的发生发展。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。通过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。蛋白表达采用 Western 印迹法分析。细胞增殖、迁移和侵袭通过(4-5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑(MTT)、5-乙炔基-2 '-脱氧尿苷(EdU)、transwell 和伤口愈合试验进行分析。circ_0000099/miR-223-3p/CTGF之间的关系通过双荧光素酶报告基因和RNA结合蛋白免疫沉淀(RIP)实验进行了验证:结果:TGF-β2 处理促进了 SRA01/04 细胞的增殖、侵袭、迁移和 EMT。敲除circ_0000099可抑制TGF-β2诱导的SRA01/04细胞增殖、侵袭、迁移和EMT。miR-223-3p被确定为circ_0000099的靶标,miR-223-3p抑制剂可部分消除circ_0000099沉默对TGF-β2诱导的SRA01/04细胞紊乱的抑制作用。MiR-223-3p 直接靶向 CTGF。敲除CTGF可抑制TGF-β2-诱导的SRA01/04细胞损伤。Circ_0000099可通过靶向miR-223-3p调节CTGF的表达:沉默Circ_0000099可通过miR-223-3p/CTGF轴缓解TGF-2诱导的SRA01/04细胞损伤,为预防和治疗PCO提供新途径。
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来源期刊
Current Eye Research
Current Eye Research 医学-眼科学
CiteScore
4.60
自引率
0.00%
发文量
163
审稿时长
12 months
期刊介绍: The principal aim of Current Eye Research is to provide rapid publication of full papers, short communications and mini-reviews, all high quality. Current Eye Research publishes articles encompassing all the areas of eye research. Subject areas include the following: clinical research, anatomy, physiology, biophysics, biochemistry, pharmacology, developmental biology, microbiology and immunology.
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