Department of Urology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China.
Department of Urology, Jiangxi Province Hospital of Integrated Chinese and Western Medicine, Nanchang, Jiangxi, China.
J Gene Med. 2022 Nov;24(11):e3418. doi: 10.1002/jgm.3418. Epub 2022 Oct 9.
Renal cell carcinoma (RCC) is a common lethal urological malignancy. Circular RNAs are assumed to play important roles in cancer development. The objective of the present study was to investigate the role and action mechanism of circ_0008717 in RCC.
The expression of circ_0008717, miR-217 and F-box protein 17 (FBXO17) mRNA was detected by a real-time quantitative polymerase chain reaction. Cell proliferation was examined using a cell counting kit-8 assay and an 5-ethynyl-2'-deoxyuridine assay. Cell apoptosis was assessed by a flow cytometry assay. Cell migration and cell invasion were investigated using a transwell assay. Glycolysis progression was assessed according to the levels of glucose uptake and lactate production. The expression of glycolysis-related proteins and FBXO17 protein was quantified by western blotting. The targets were analyzed by the bioinformatics tools (starBase and circinteractome) and validated by a dual-luciferase reporter assay, RNA pull-down assay and RNA immunoprecipitation assay. A xenograft model was established to monitor the role of circ_0008717 in vivo.
Circ_0008717 was upregulated in RCC tissues and cells. Silencing circ_0008717 suppressed RCC cell proliferation, migration, invasion and glycolysis but promoted cell apoptosis. MiR-217 was a target of circ_0008717 and bound to the FBXO17 3' untranslated region. The expression of FBXO17 was positively regulated by circ_0008717 but impaired by miR-217 reintroduction. The inhibitory effects of circ_0008717 knockdown on RCC cell malignant behaviors were reversed by miR-217 inhibition or FBXO17 overexpression. Circ_0008717 knockdown inhibited tumor growth in vivo by regulating miR-217 and FBXO17.
Circ_0008717 aggravated the progression of RCC by activating FBXO17 through targeting miR-217, which provided a novel mechanism for circ_0008717 to participate in RCC progression.
肾细胞癌（RCC）是一种常见的致命性泌尿系统恶性肿瘤。环状 RNA 被认为在癌症发展中发挥重要作用。本研究旨在探讨 circ_0008717 在 RCC 中的作用和作用机制。
通过实时定量聚合酶链反应检测 circ_0008717、miR-217 和 F-box 蛋白 17（FBXO17）mRNA 的表达。使用细胞计数试剂盒-8 测定法和 5-乙炔基-2'-脱氧尿苷测定法检测细胞增殖。通过流式细胞术检测细胞凋亡。通过 Transwell 测定法检测细胞迁移和细胞侵袭。根据葡萄糖摄取和乳酸产生水平评估糖酵解进展。通过 Western blot 定量测定糖酵解相关蛋白和 FBXO17 蛋白的表达。通过生物信息学工具（starBase 和 circinteractome）分析靶标，并通过双荧光素酶报告基因测定、RNA 下拉测定和 RNA 免疫沉淀测定进行验证。建立异种移植模型以监测 circ_0008717 在体内的作用。
circ_0008717 在 RCC 组织和细胞中上调。沉默 circ_0008717 抑制 RCC 细胞增殖、迁移、侵袭和糖酵解，但促进细胞凋亡。miR-217 是 circ_0008717 的靶标，并与 FBXO17 3'非翻译区结合。FBXO17 的表达受 circ_0008717 的正向调节，但受 miR-217 再导入的抑制。circ_0008717 敲低对 RCC 细胞恶性行为的抑制作用通过 miR-217 抑制或 FBXO17 过表达而逆转。circ_0008717 敲低通过调节 miR-217 和 FBXO17 抑制体内肿瘤生长。
circ_0008717 通过靶向 miR-217 激活 FBXO17 加重 RCC 的进展，为 circ_0008717 参与 RCC 进展提供了新的机制。
