Department of Urology, Section 4, People's Hospital of Hunan Province, No.89, Guhan Road, Furong District, Changsha, 410002, Hunan, China.
Biochem Genet. 2022 Aug;60(4):1346-1361. doi: 10.1007/s10528-021-10160-w. Epub 2022 Jan 7.
Prostate cancer (PCa) is one of the most common and deadly cancers in men. The dysregulated circular RNAs (circRNAs) are involved in the development of various cancers, including PCa. The purpose of this study was to construct a circRNA-microRNA (miRNA)-mRNA network to explain the function of circ_0044516 in PCa. The expression analysis of circ_0044516, miR-330-5p, and sterile alpha and TIR motif-containing 1 (SARM1) was performed using real-time quantitative polymerase chain reaction, and the protein level of SARM1 was detected by western blot. The interaction between miR-330-5p and circ_0044516 or SARM1 obtained by bioinformatics prediction was verified by dual-luciferase reporter assay or RNA immunoprecipitation assay. For functional studies, cell proliferation was assessed by cell viability and colony formation ability using cell counting kit-8 assay and colony formation assay. Cell migration and invasion were studied using transwell assay. Cell apoptosis and cell cycle were investigated using flow cytometry assay. The tumorigenicity of circ_0044516 was tested by animal study. Circ_0044516 and SARM1 were highly expressed, while miR-330-5p was rarely expressed in PCa tissues and cells. Circ_0044516 acted as a miR-330-5p sponge to block miR-330-5p expression, and circ_0044516 knockdown suppressed PCa cell proliferation, migration, invasion, and cycle progression by enriching miR-330-5p. SARM1 was a target of miR-330-5p, and miR-330-5p restoration also inhibited PCa cell proliferation, migration, invasion, and cycle progression by degrading SARM1. Moreover, circ_0044516 deficiency blocked tumor growth in vivo by regulating the miR-330-5p/SARM1 axis. Circ_0044516 as a miR-330-5p sponge increases SARM1 expression, thus promoting the malignant development of PCa.
前列腺癌(PCa)是男性最常见和最致命的癌症之一。失调的环状 RNA(circRNAs)参与了多种癌症的发展,包括 PCa。本研究的目的是构建一个 circRNA- microRNA(miRNA)-mRNA 网络,以解释 circ_0044516 在 PCa 中的功能。使用实时定量聚合酶链反应分析 circ_0044516、miR-330-5p 和无菌α和 TIR 基序包含 1(SARM1)的表达,并通过 Western blot 检测 SARM1 的蛋白水平。通过生物信息学预测验证 miR-330-5p 与 circ_0044516 或 SARM1 之间的相互作用,通过双荧光素酶报告基因测定或 RNA 免疫沉淀测定。为了进行功能研究,通过细胞计数试剂盒-8 测定和集落形成测定评估细胞增殖,以评估细胞活力和集落形成能力。使用 Transwell 测定研究细胞迁移和侵袭。通过流式细胞术测定研究细胞凋亡和细胞周期。通过动物研究测试 circ_0044516 的致瘤性。circ_0044516 和 SARM1 在 PCa 组织和细胞中高表达,而 miR-330-5p 则很少表达。circ_0044516 作为 miR-330-5p 的海绵,阻断 miR-330-5p 的表达,circ_0044516 敲低通过富集 miR-330-5p 抑制 PCa 细胞增殖、迁移、侵袭和周期进展。SARM1 是 miR-330-5p 的靶标,miR-330-5p 恢复通过降解 SARM1 也抑制 PCa 细胞增殖、迁移、侵袭和周期进展。此外,circ_0044516 缺乏通过调节 miR-330-5p/SARM1 轴在体内阻断肿瘤生长。circ_0044516 作为 miR-330-5p 的海绵增加 SARM1 的表达,从而促进 PCa 的恶性发展。