Xu Le, Long Hua, Zhou Bo, Jiang Haibo, Cai Mingfang
Department of Ophthalmology, Suizhou Hospital, Hubei University of Medicine, Suizhou City, Hubei Province, China.
Curr Eye Res. 2021 Nov;46(11):1751-1761. doi: 10.1080/02713683.2021.1927110. Epub 2021 May 14.
: Circular RNAs (circRNAs) are essential regulators in tumorigenesis and development. In this study, we focused on the functions of circRNA muskelin 1 (circMKLN1) in retinoblastoma (RB) progression.: Quantitative real-time polymerase chain reaction (qRT-PCR) assay was conducted to determine the levels of circMKLN1, microRNA-425-5p (miR-425-5p) and programmed cell death 4 (PDCD4). The characteristic of circMKLN1 was analyzed using RNase R assay. Cell Counting Kit-8 (CCK-8) assay and colony formation assay were employed to explore cell proliferation ability. The transwell assay was utilized for cell migration and invasion. A Western blot assay was performed for protein levels. The dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were conducted to demonstrate the relationships among circMKLN1, miR-425-5p and PDCD4. Murine xenograft model assay was adopted to investigate the role of circMKLN1 .: CircMKLN1 was downregulated in RB tissues and cells. High levels of circMKLN1 were related to a favorable outcome of RB patients. CircMKLN1 was resistant to RNase R digestion and circMKLN1 overexpression repressed RB cell proliferation, migration and invasion . MiR-425-5p was identified as the target of circMKLN1 and miR-425-5p elevation reversed the effects of circMKLN1 overexpression on RB cell malignant behaviors. Furthermore, as the target gene of miR-425-5p, PDCD4 silencing could ameliorate the suppressive roles of circMKLN1 in RB cell growth and metastasis. Additionally, circMKLN1 overexpression hampered tumor growth .: CircMKLN1 overexpression decelerated the progression of RB through sponging miR-425-5p and elevating PDCD4.
环状RNA(circRNAs)是肿瘤发生和发展过程中的重要调节因子。在本研究中,我们聚焦于环状RNA肌肉萎缩相关蛋白1(circMKLN1)在视网膜母细胞瘤(RB)进展中的功能。
进行定量实时聚合酶链反应(qRT-PCR)检测以确定circMKLN1、微小RNA-425-5p(miR-425-5p)和程序性细胞死亡4(PDCD4)的水平。使用核糖核酸酶R检测分析circMKLN1的特征。采用细胞计数试剂盒-8(CCK-8)检测和集落形成检测来探究细胞增殖能力。运用Transwell检测评估细胞迁移和侵袭能力。进行蛋白质免疫印迹检测以分析蛋白水平。采用双荧光素酶报告基因检测和RNA免疫沉淀(RIP)检测来证明circMKLN1、miR-425-5p和PDCD4之间的关系。采用小鼠异种移植模型检测来研究circMKLN1的作用。
circMKLN1在RB组织和细胞中表达下调。circMKLN1高表达与RB患者的良好预后相关。circMKLN1对核糖核酸酶R消化具有抗性,circMKLN1过表达可抑制RB细胞的增殖、迁移和侵袭。miR-425-5p被鉴定为circMKLN1的靶标,miR-425-5p表达上调可逆转circMKLN1过表达对RB细胞恶性行为的影响。此外,作为miR-425-5p的靶基因,PDCD4沉默可改善circMKLN1对RB细胞生长和转移的抑制作用。此外,circMKLN1过表达可抑制肿瘤生长。
circMKLN1过表达通过吸附miR-425-5p和上调PDCD4来减缓RB的进展。