Luo Dongdong, Luo Aiping, Ye Ganwei, Li Dan, Hu Su, Zhao Hailin, Peng Biao
Department of Neurosurgery, Guangzhou Institute of Cancer Research, the Affiliated Cancer Hospital, Guangzhou Medical University, No. 78 Hengzhigang Road, Yuexiu District, Guangzhou, 510095, Guangdong, China.
Department of Radiology, Guangzhou Institute of Cancer Research, the Affiliated Cancer Hospital, Guangzhou Medical University, No. 78 Hengzhigang Road, Yuexiu District, Guangzhou, 510095, Guangdong, China.
J Radiat Res. 2024 Dec 3;65(6):752-764. doi: 10.1093/jrr/rrae064.
Deregulation of circular RNAs (circRNAs) is frequent in human glioma. Although circRNA ATPase phospholipid transporting 8B4 (circATP8B4) is highly expressed in glioma, its precise action in glioma development is still not fully understood. The relationship of microRNA (miR)-31-5p and circATP8B4 or nestin (NES) was predicted by bioinformatic analysis and confirmed by RNA pull-down and Dual-luciferase reporter assays. CircATP8B4, miR-31-5p and NES were quantified by qRT-PCR or western blot. Cell functional behaviors were assessed by EdU, wound-healing and transwell invasion assays. Xenograft model experiments were performed to define circATP8B4's activity in vivo. CircATP8B4, a true circular transcript, was upregulated in human glioma. CircATP8B4 downregulation weakened glioma cell growth, motility, and invasion and facilitated radiosensitivity. Mechanistically, circATP8B4 and NES 3'UTR harbored a shared miR-31-5p pairing site, and circATP8B4 involved the post-transcriptional NES regulation by functioning as a competing endogenous RNA (ceRNA). Furthermore, the miR-31-5p/NES axis participated in circATP8B4's activity in glioma cell proliferation, motility, invasion and radiosensitivity. Additionally, circATP8B4 loss diminished tumor growth and enhanced the anticancer effect of radiotherapy in vivo. We have uncovered an uncharacterized ceRNA cascade, circATP8B4/miR-31-5p/NES axis, underlying glioma development and radiosensitivity. Targeting the ceRNA crosstalk may have potential to improve the outcome of glioma patients.
环状RNA（circRNAs）失调在人类胶质瘤中很常见。尽管环状RNA三磷酸腺苷磷脂转运体8B4（circATP8B4）在胶质瘤中高表达，但其在胶质瘤发生发展中的具体作用仍未完全明确。通过生物信息学分析预测了微小RNA（miR）-31-5p与circATP8B4或巢蛋白（NES）的关系，并通过RNA下拉实验和双荧光素酶报告基因检测进行了验证。通过qRT-PCR或蛋白质免疫印迹法对circATP8B4、miR-31-5p和NES进行定量分析。通过EdU实验、伤口愈合实验和Transwell侵袭实验评估细胞功能行为。进行异种移植模型实验以确定circATP8B4在体内的活性。CircATP8B4是一种真正的环状转录本，在人类胶质瘤中上调。CircATP8B4下调可减弱胶质瘤细胞的生长、运动和侵袭能力，并提高放射敏感性。机制上，circATP8B4和NES的3'UTR具有共同的miR-31-5p配对位点，circATP8B4作为竞争性内源性RNA（ceRNA）参与NES的转录后调控。此外，miR-31-5p/NES轴参与了circATP8B4在胶质瘤细胞增殖、运动、侵袭和放射敏感性方面的活性。此外，circATP8B4缺失可减少肿瘤生长并增强体内放疗的抗癌效果。我们发现了一种未被描述的ceRNA级联反应，即circATP8B4/miR-31-5p/NES轴，它是胶质瘤发生发展和放射敏感性的基础。靶向ceRNA相互作用可能有改善胶质瘤患者预后的潜力。
