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环状MKLN1通过调节miR-425-5p/PDCD4轴抑制人视网膜母细胞瘤的进展。

CircMKLN1 Suppresses the Progression of Human Retinoblastoma by Modulation of miR-425-5p/PDCD4 Axis.

作者信息

Xu Le, Long Hua, Zhou Bo, Jiang Haibo, Cai Mingfang

机构信息

Department of Ophthalmology, Suizhou Hospital, Hubei University of Medicine, Suizhou City, Hubei Province, China.

出版信息

Curr Eye Res. 2021 Nov;46(11):1751-1761. doi: 10.1080/02713683.2021.1927110. Epub 2021 May 14.

Abstract

: 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.

摘要

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。

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