Department of Urology WardIII, Huangshi Central Hospital, Affiliated Hospital of Hubei Polytechnic University, Edong Healthcare Group, No.293, Feiyun Street, Huangshi, 435000, Hubei Province, China.
Hubei Key Laboratory of Kidney Disease Pathogenesis and Intervention, Wuhan, China.
Biochem Genet. 2023 Oct;61(5):1987-2003. doi: 10.1007/s10528-022-10328-y. Epub 2023 Mar 13.
This study aimed to investigate more detailed functions of circ_0005276 in prostate cancer (PCa) and provide a novel mechanism for circ_0005276 action. The expression of circ_0005276, microRNA-128-3p (miR-128-3p) and DEP domain containing 1B (DEPDC1B) was detected by quantitative real-time PCR. In functional assays, cell proliferation was determined by CCK-8 assay and EdU assay. Cell migration and invasion were determined by transwell assay. The ability of angiogenesis was determined by tube formation assay. Cell apoptosis was determined by flow cytometry assay. The potential binding relationship between miR-128-3p and circ_0005276 or DEPDC1B was ascertained by dual-luciferase reporter assay and RIP assay. Mouse models were used to verify the role of circ_0005276 in vivo. The upregulation of circ_0005276 was determined in PCa tissues and cells. Circ_0005276 knockdown inhibited proliferation, migration, invasion and angiogenesis in PCa cells, and circ_0005276 knockdown also blocks tumor growth in vivo. Mechanism analysis discovered that miR-128-3p was a target of circ_0005276, and miR-128-3p inhibition recovered circ_0005276 knockdown-inhibited proliferation, migration, invasion and angiogenesis. In addition, DEPDC1B was a target of miR-128-3p, and miR-128-3p restoration-inhibited proliferation, migration, invasion and angiogenesis were rescued by DEPDC1B overexpression. Circ_0005276 might promote the development of PCa by activating the expression of DEPDC1B via targeting miR-128-3p.
本研究旨在探究 circ_0005276 在前列腺癌（PCa）中的更详细功能，并为 circ_0005276 的作用提供新的机制。通过定量实时 PCR 检测 circ_0005276、microRNA-128-3p（miR-128-3p）和 DEP 结构域包含 1B（DEPDC1B）的表达。在功能测定中，通过 CCK-8 测定和 EdU 测定来确定细胞增殖。通过 Transwell 测定来确定细胞迁移和侵袭。通过管形成测定来确定血管生成能力。通过流式细胞术测定来确定细胞凋亡。通过双荧光素酶报告基因测定和 RIP 测定来确定 miR-128-3p 与 circ_0005276 或 DEPDC1B 的潜在结合关系。使用小鼠模型验证 circ_0005276 在体内的作用。在 PCa 组织和细胞中确定 circ_0005276 的上调。circ_0005276 敲低抑制 PCa 细胞的增殖、迁移、侵袭和血管生成，并且 circ_0005276 敲低也阻止体内肿瘤生长。机制分析发现 miR-128-3p 是 circ_0005276 的靶标，并且 miR-128-3p 抑制恢复了 circ_0005276 敲低抑制的增殖、迁移、侵袭和血管生成。此外，DEPDC1B 是 miR-128-3p 的靶标，并且 miR-128-3p 恢复抑制的增殖、迁移、侵袭和血管生成通过过表达 DEPDC1B 得到挽救。circ_0005276 可能通过靶向 miR-128-3p 激活 DEPDC1B 的表达来促进 PCa 的发展。
