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环状 RNA_CCNB2 通过 miR-30b-5p/KIF18A 轴抑制自噬使前列腺癌对放射敏感。

Knockdown of Circ_CCNB2 Sensitizes Prostate Cancer to Radiation Through Repressing Autophagy by the miR-30b-5p/KIF18A Axis.

机构信息

Department of Urology Surgery, the Second Affiliated Hospital of Fujian Medical University, Quanzhou, China.

出版信息

Cancer Biother Radiopharm. 2022 Aug;37(6):480-493. doi: 10.1089/cbr.2019.3538. Epub 2020 Jul 23.

Abstract

Circular RNAs (circRNAs) have recently emerged as crucial regulatory molecules in prostate cancer (PCa), but few researches focus on the effects of circRNAs on PCa radiosensitivity. The issue will be addressed in this study using circRNA Cyclin B2 (circ_CCNB2) as an object. All RNA and protein levels were severally examined using quantitative real-time polymerase chain reaction and Western blot. Colony formation assay and flow cytometry were implemented for detecting cell colony capacity and apoptotic cells, respectively. Cellular migration and invasion abilities were evaluated by transwell assay. The combination between potential target molecules was analyzed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. The effect of circ_CCNB2 on PCa radiosensitivity was explored using xenograft models in mice. Circ_CCNB2 was upregulated in irradiation-resistant PCa tissues and cells. Circ_CCNB2 knockdown had promoted effect on the radiosensitivity of irradiation-resistant PCa cells by inhibiting autophagy. Besides, circ_CCNB2 could directly sponge miR-30b-5p, and the promotion of circ_CCNB2 knockdown on PCa radiosensitivity was achieved by elevating miR-30b-5p. MiR-30b-5p enhanced the radiosensitivity of irradiation-resistant PCa cells through repressing the expression of its target kinesin family member 18A (KIF18A). Furthermore, circ_CCNB2 regulated the KIF18A level through targeting miR-30b-5p. Circ_CCNB2 downregulation facilitated PCa radiosensitivity through inhibiting autophagy by miR-30b-5p/KIF18A. In this study, knockdown of circ_CCNB2 was shown to promote PCa radiosensitivity through autophagy repression by miR-30b-5p/KIF18A axis, developing a molecular resistance mechanism of PCa radiotherapy and a feasible strategy to increase radiosensitivity.

摘要

环状 RNA(circRNAs)最近被发现为前列腺癌(PCa)中的重要调控分子,但很少有研究关注 circRNAs 对 PCa 放射敏感性的影响。本研究以环状 RNA 细胞周期素 B2(circ_CCNB2)为研究对象,探讨了这一问题。分别采用实时定量聚合酶链反应和 Western blot 检测所有 RNA 和蛋白质水平。采用集落形成实验和流式细胞术检测细胞集落形成能力和凋亡细胞。通过 Transwell 实验评估细胞迁移和侵袭能力。通过双荧光素酶报告和 RNA 免疫沉淀(RIP)实验分析潜在靶分子之间的结合。在小鼠异种移植模型中探讨了 circ_CCNB2 对 PCa 放射敏感性的影响。在辐射抗性 PCa 组织和细胞中,circ_CCNB2 上调。circ_CCNB2 敲低通过抑制自噬促进辐射抗性 PCa 细胞的放射敏感性。此外,circ_CCNB2 可以直接吸附 miR-30b-5p,circ_CCNB2 敲低通过提高 miR-30b-5p 来实现对 PCa 放射敏感性的促进。miR-30b-5p 通过抑制其靶基因驱动蛋白家族成员 18A(KIF18A)的表达,增强了辐射抗性 PCa 细胞的放射敏感性。此外,circ_CCNB2 通过靶向 miR-30b-5p 调节 KIF18A 水平。circ_CCNB2 下调通过 miR-30b-5p/KIF18A 轴抑制自噬,促进 PCa 放射敏感性。在本研究中,circ_CCNB2 的敲低通过 miR-30b-5p/KIF18A 轴抑制自噬,促进了 PCa 的放射敏感性,为 PCa 放射治疗的分子抵抗机制和提高放射敏感性的可行策略提供了依据。

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