环状 RNA circ_0000034 通过抑制 miR-361-3p 上调 STX17 水平促进人视网膜母细胞瘤的发展。
Circular RNA circ_0000034 upregulates STX17 level to promote human retinoblastoma development via inhibiting miR-361-3p.
机构信息
Department of Ophthalmology, Daping Hospital, Army Medical Center of PLA, Chongqing, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Dec;24(23):12080-12092. doi: 10.26355/eurrev_202012_23997.
OBJECTIVE
Retinoblastoma (RB) is a common intraocular tumor of infancy and childhood. Circular RNAs (circRNAs) are related to the development of RB. The purpose of this research was to reveal the functional mechanism of circRNA circ_0000034 in RB.
MATERIALS AND METHODS
Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and Western blot were applied to determine the levels of genes. MTT assay and flow cytometry were employed to assess cell proliferation and apoptosis rate, respectively. Furthermore, cell migratory and invasive abilities were measured using the transwell assay. Mouse xenograft was conducted to analyze the effect of circ_0000034 on tumor growth in vivo. Besides, the interaction between miR-361-3p and circ_0000034 or syntaxin 17 (STX17) was predicted by starBase, and then, confirmed by the Dual-Luciferase reporter assay and RNA immunoprecipitation (RIP) assay.
RESULTS
The levels of circ_0000034 and STX17 were increased and miR-361-3p level was decreased in RB tissues and cells. Circ_0000034 knockdown suppressed cell proliferation, migration, invasion, autophagy, and tumor growth, and induced apoptosis in RB. Circ_0000034 targeted miR-361-3p and miR-361-3p bound to STX17. Circ_0000034 overexpression and miR-361-3p knockdown reversed the effect of miR-361-3p upregulation and STX17 depletion on the growth of RB cells, respectively. Besides, circ_0000034 elevated STX17 level by repressing miR-361-3p expression.
CONCLUSIONS
We demonstrated that circ_0000034 knockdown suppressed the development of RB by the modulation of miR-361-3p/STX17 axis. Our findings provided a theoretical basis for the treatment of RB.
目的
视网膜母细胞瘤（RB）是一种常见的婴幼儿眼内肿瘤。环状 RNA（circRNA）与 RB 的发生发展有关。本研究旨在揭示环状 RNA circ_0000034 在 RB 中的功能机制。
材料和方法
采用实时定量聚合酶链反应（qRT-PCR）和 Western blot 检测基因水平。分别采用 MTT 法和流式细胞术检测细胞增殖和凋亡率，采用 Transwell 检测细胞迁移和侵袭能力。通过小鼠异种移植分析 circ_0000034 对体内肿瘤生长的影响。此外，通过 starBase 预测 miR-361-3p 与 circ_0000034 或 syntaxin 17（STX17）的相互作用，然后通过双荧光素酶报告基因检测和 RNA 免疫沉淀（RIP）实验验证。
结果
RB 组织和细胞中 circ_0000034 和 STX17 水平升高，miR-361-3p 水平降低。circ_0000034 敲低抑制 RB 细胞的增殖、迁移、侵袭、自噬和肿瘤生长，并诱导细胞凋亡。Circ_0000034 靶向 miR-361-3p，miR-361-3p 与 STX17 结合。Circ_0000034 过表达和 miR-361-3p 敲低分别逆转了 miR-361-3p 上调和 STX17 耗竭对 RB 细胞生长的影响。此外，circ_0000034 通过抑制 miR-361-3p 的表达来升高 STX17 水平。
结论
我们证实，circ_0000034 敲低通过调节 miR-361-3p/STX17 轴抑制 RB 的发展。我们的研究结果为 RB 的治疗提供了理论依据。
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