Ophthalmology Department, Baicheng Central Hospital, No.111 Zhongxing West Road, Taobei District, Baicheng, Jilin, 137000, China.
Ophthalmology Department, Changchun Bokangming Eye Hospital, Changchun, Jilin, 130000, China.
Mol Biotechnol. 2023 Oct;65(10):1608-1618. doi: 10.1007/s12033-023-00660-y. Epub 2023 Jan 30.
Increasing evidences indicate the crucial role of circRNAs in tumorigenesis, but little is understood about their molecular mechanism in retinoblastoma (RB). This paper was designed for exploring the circ_0119412 function in cases with RB and the potential mechanism. RT-qPCR was utilized to ascertain circ_0119412 and miR-186-5p levels in RB tissues and cells, and western blotting was used to quantify ELK4 in RB cells. In addition, CCK-8 and scratch assays were carried out for evaluation of cell proliferation and migration, respectively. Apoptosis-related proteins levels (Bax and Bcl-2) were measure by western blotting. Tumor growth in vivo was detected utilizing xenograft tumor experiment. The targeting relationship between circ_0119412, miR-186-5p, and ELK4 was validated using a dual-luciferase reporter assay and an RNA immunoprecipitation (RIP) assay. In RB tissues and cells, Circ_0119412 and ELK4 expression were upregulated, while miR-186-5p expression was downregulated. In vitro assay revealed that downregulating circ_0119412 accelerated the cell apoptosis of RB cells and slowed down their migration and proliferation, and the in vivo assay indicated that circ_0119412 downregulation reduced the weight and volume of tumor in nude mice. In addition, miR-186-5p interference promoted the malignant behavior of RB cells, while ELK4 silencing showed an opposite trend. Mechanically, circ_0119412 can promote RB malignant phenotypes via miR-186-5p/ELK4 axis. Circ_0119412 was found to be upregulated in RB, and could accelerate the progression of RB via the miR-186-5p/ELK4 axis, indicating circ_0119412 may serve a promising clinical therapeutic target of RB.
越来越多的证据表明 circRNAs 在肿瘤发生中起着关键作用，但人们对其在视网膜母细胞瘤 (RB) 中的分子机制知之甚少。本文旨在探讨 circ_0119412 在 RB 中的作用及其潜在机制。利用 RT-qPCR 检测 RB 组织和细胞中 circ_0119412 和 miR-186-5p 的水平，利用 Western blot 检测 RB 细胞中 ELK4 的含量。此外，分别通过 CCK-8 和划痕实验评估细胞增殖和迁移能力。通过 Western blot 检测细胞凋亡相关蛋白 (Bax 和 Bcl-2) 水平。利用异种移植瘤实验检测体内肿瘤生长情况。利用双荧光素酶报告基因实验和 RNA 免疫沉淀 (RIP) 实验验证 circ_0119412、miR-186-5p 和 ELK4 之间的靶向关系。在 RB 组织和细胞中，circ_0119412 和 ELK4 的表达上调，而 miR-186-5p 的表达下调。体外实验表明，下调 circ_0119412 可促进 RB 细胞凋亡，抑制其迁移和增殖，体内实验表明，下调 circ_0119412 可减少裸鼠肿瘤的重量和体积。此外，miR-186-5p 干扰促进了 RB 细胞的恶性行为，而 ELK4 沉默则表现出相反的趋势。机制上，circ_0119412 通过 miR-186-5p/ELK4 轴促进 RB 恶性表型。研究发现 circ_0119412 在 RB 中上调，并可通过 miR-186-5p/ELK4 轴加速 RB 的进展，表明 circ_0119412 可能成为 RB 有前途的临床治疗靶点。
