Department of Neurosurgery, Weifang Yidu Central Hospital, Qingzhou, 262500, Shandong, China.
Department of Pediatrics, Central Hospital of Linyi, Linyi, 276400, Shandong, China.
Mol Biotechnol. 2022 Sep;64(9):1022-1033. doi: 10.1007/s12033-022-00466-4. Epub 2022 Mar 29.
Circular RNAs (circRNAs) are reported to participate in the development of diverse human malignancies. This work investigated the mechanism of circSKA3 in modulating medulloblastoma progression. A total of 15 cases of medulloblastoma were collected in this work. Daoy cells were used to construct cell models. The expression level of circSKA3, microRNA-520 h (miR-520 h), and cyclin-dependent kinase 6 (CDK6) mRNA in tissues or cells was examined by quantitative real-time polymerase chain reaction (qRT-PCR). Western blot was employed to detect CDK6 protein expression. CCK-8 experiment, Transwell assay, and flow cytometry were applied to detect the regulatory effects of circSKA3 on cell proliferation, migration, invasion, and cell cycle. Dual-luciferase reporter gene experiment was executed to determine the relationship between circSKA3 and miR-520 h, and between miR-520 h and CDK6. circSKA3 was remarkably up-modulated in medulloblastoma tissues. CircSKA3 depletion markedly suppressed Daoy cell viability, migration, invasion, and cell cycle progression. CircSKA3 overexpression induced the opposite effects. circSKA3 could decoyed miR-520 h, which targeted the 3' UTR of CDK6. circSKA3 expression in medulloblastoma tissues was negatively correlated with miR-520 h expression and positively correlated with CDK6 expression. "Rescue" experiments revealed that miR-520 h down-modulation or CDK6 overexpression remarkably counteracted the inhibitory effect of circSKA3 knockdown on Daoy cells. circSKA3 facilitates medulloblastoma progression through miR-520 h/CDK6.
环状 RNA（circRNAs）被报道参与多种人类恶性肿瘤的发生发展。本研究旨在探讨 circSKA3 在调节髓母细胞瘤进展中的作用机制。本研究共收集了 15 例髓母细胞瘤病例。使用 Daoy 细胞构建细胞模型。通过实时定量聚合酶链反应（qRT-PCR）检测组织或细胞中 circSKA3、microRNA-520h（miR-520h）和细胞周期蛋白依赖性激酶 6（CDK6）mRNA 的表达水平。采用 Western blot 检测 CDK6 蛋白表达。通过 CCK-8 实验、Transwell 实验和流式细胞术检测 circSKA3 对细胞增殖、迁移、侵袭和细胞周期的调控作用。双荧光素酶报告基因实验确定 circSKA3 与 miR-520h 之间以及 miR-520h 与 CDK6 之间的关系。髓母细胞瘤组织中 circSKA3 表达显著上调。circSKA3 缺失显著抑制 Daoy 细胞活力、迁移、侵袭和细胞周期进程。circSKA3 过表达则诱导相反的效果。circSKA3 可以与 miR-520h 结合，而 miR-520h 可以靶向 CDK6 的 3'UTR。髓母细胞瘤组织中 circSKA3 的表达与 miR-520h 的表达呈负相关，与 CDK6 的表达呈正相关。“挽救”实验表明，miR-520h 下调或 CDK6 过表达显著抵消了 circSKA3 敲低对 Daoy 细胞的抑制作用。circSKA3 通过 miR-520h/CDK6 促进髓母细胞瘤的进展。
