外泌体circRNA_104948通过调控miR-29b-3p和DNMT3B/MTSS1信号增强胶质瘤的进展

Shoudan Zhang, Ning Guan, Xin Mao, Jingwen Cui, Xin Sui, Wenshi Guo
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引用次数: 5

摘要

胶质瘤是中枢神经系统常见的一种恶性肿瘤。胶质瘤的发病机制十分复杂,其潜在机制在很大程度上仍不为人所知。我们的研究从患者样本中提取了外泌体,并通过透射电子显微镜对分离出的外泌体进行了确认。利用 RT-qPCR 测定了 circRNA_104948、miR-29b-3p 和 DNMT3B 的表达。细胞增殖活性采用 CCK-8 法检测。细胞凋亡率通过流式细胞术进行评估。增殖或细胞凋亡标志物的表达水平通过 Western 印迹法进行测定。我们的数据表明,circRNA_104948在胶质瘤患者和胶质瘤细胞系的血浆外泌体/组织样本中上调。此外,用外泌体circRNA_104948处理正常星形胶质细胞后,细胞增殖增强,凋亡被抑制,而sh-circRNA_104948能逆转这些影响。此外,miR-29b-3p 是 circRNA_104948 的一个新靶点,而 DNMT3B 是 miR-29b-3p 的推定下游分子。circRNA_104948可通过miR-29b-3p/DNMT3B/MTSS1信号转导调控星形胶质细胞的增殖/凋亡,miR-29b-3p模拟物可逆转胶质瘤-Exo诱导的生物学行为变化;敲除DNMT3B可减轻miR-29b-3p抑制剂导致的细胞生长上调;过表达DNMT3B可取消miR-29b-3p模拟物诱导的效应。我们的研究结果揭示了circRNA_104948在胶质瘤发病过程中的重要作用,circRNA_104948/miR-29b-3p/MTSS1/DNMT3B通路可能是胶质瘤患者靶向治疗的潜在候选通路。
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Exosomal circRNA_104948 Enhances the Progression of Glioma by Regulating miR-29b-3p and DNMT3B/MTSS1 Signaling.

Glioma is a common type of malignancy in the central nervous system. The pathogenesis of glioma is complex and the underlying mechanisms remain largely unknown. In our study, exosomes were exacted from patient samples, and the isolated exosomes were confirmed by transmission electron microscope. The expression of circRNA_104948, miR-29b-3p and DNMT3B were determined using RT-qPCR. Proliferative activity of cell was examined using CCK-8 assay. Cell apoptotic rate was evaluated by flow cytometry. The expression levels of proliferation or apop-tosis markers were determined using western blotting. Our data suggested that circRNA_104948 was upregulated in plasma exosomes/tissue samples of glioma patients and glioma cell lines. Furthermore, cell proliferation was enhanced and apoptosis was suppressed in normal astrocytes treated with exosomal circRNA_104948, and the effects were reversed by sh-circRNA_104948. In addition, miR-29b-3p is a novel target of circRNA_104948, and DNMT3B is a putative downstream molecule of miR-29b-3p. circRNA_104948 could regulate the proliferation/apoptosis of astrocytes through miR-29b-3p/DNMT3B/MTSS1 signaling, and the biological behavior changes induced by glioma-Exo were reversed by miR-29b-3p mimics; upregulated cell growth caused by miR-29b-3p inhibitors was abrogated by the knockdown of DNMT3B; the effects induced by miR-29b-3p mimics were abolished by the overexpression of DNMT3B. Our findings revealed the important roles of circRNA_104948 on the development of glioma, and circRNA_104948/miR-29b-3p/MTSS1/DNMT3B pathway may be a potential candidate for the target therapy of glioma patients.

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