miR-423-5p contributes to a malignant phenotype and temozolomide chemoresistance in glioblastomas
miR-423-5p contributes to a malignant phenotype and temozolomide chemoresistance in glioblastomas
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miR-423-5p 导致胶质母细胞瘤的恶性表型和替莫唑胺化疗耐药
DOI：
10.1093/neuonc/now129
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发表时间：
2017-01-01
期刊：
影响因子：
15.9
通讯作者：
You, Yongping
中科院分区：
文献类型：
--
作者：
Li, Shouwei;Zeng, Ailiang;You, Yongping
背景神经胶质瘤是一种基于遗传异常的肿瘤，其预后很差。MicroRNAs(MiRNAs)被认为是胶质瘤组织中基因表达的重要介质。方法采用实时定量聚合酶链式反应(Real-Time PCR)技术检测microRNA-423-5p(miR-423-5p)在人脑胶质瘤组织和正常脑组织中的表达。采用细胞凋亡率、细胞周期、细胞增殖、免疫染色、Transwell、体外2D和3D迁移及药敏试验等方法检测过表达miRNA-423-5p的胶质瘤细胞的表型变化。Western blotting检测生长抑制因子4(ING-4)在脑胶质瘤组织中的表达，荧光素酶报告实验证实ING-4是否是miR-423-5p的直接靶点。Western blotting鉴定miR-423-5p影响胶质瘤细胞生长的潜在信号通路。体内实验观察miR-423-5P对胶质瘤组织的致癌作用。结果我们首次报道了miR-423-5p在胶质瘤中的表达增加，并通过靶向ING-4成为潜在的肿瘤促进剂。MiR-423-5p的过度表达导致重要的信号分子p-AKT和p-ERK1/2的上调。在临床标本中，miR-423-5p的表达异常，ING-4的表达也发生了相应的变化(P=0.0207)。此外，miR-423-5p的过表达增强了胶质瘤细胞的增殖、血管生成和侵袭性。最后，miR-423-5p的过表达也加强了GBM神经球的形成，并使胶质瘤细胞对替莫唑胺(TMZ)产生耐药。结论miR-423-5p通过抑制ING-4在脑胶质瘤组织中发挥癌基因的作用，提示miR-423-5p具有治疗脑胶质瘤的潜力。
Background Gliomas are based on a genetic abnormality and present with a dismal prognosis. MicroRNAs (miRNAs) are considered to be important mediators of gene expression in glioma tissues. Methods Real-time PCR was used to analyze the expression of microRNA-423-5p (miR-423-5p) in human glioma samples and normal brain tissue. Apoptosis, cell cycle, proliferation, immunostaining, transwell, in vitro 2D and 3D migration, and chemosensitivity assays were performed to assess the phenotypic changes in glioma cells overexpressing miRNA-423-5p. Western blotting was used to determine the expression of inhibitor of growth 4 (ING-4)in glioma tissues, and a luciferase reporter assay was conducted to confirm whether ING-4 is a direct target of miR-423-5p. Western blotting was used to identify the potential signaling pathways that are affected in glioma cell growth by miR-423-5p. Xenograft tumors were examined in vivo for the carcinogenic effects of miR-423-5p in glioma tissues. Results We first reported that miR-423-5p expression was increased in gliomas and was a potential tumor promoter via targeting ING-4. The overexpression of miR-423-5p resulted in upregulation of important signaling molecules such as p-AKT and p-ERK1/2. In clinical samples, miR-423-5p was dysregulated, and a corresponding alteration in ING-4 expression was observed (P = .0207). Furthermore, the overexpression of miR-423-5p strengthened glioma cell proliferation, angiogenesis, and invasion. Finally, miR-423-5p overexpression also strengthened GBM neurosphere formation and rendered glioma cells resistant to temozolomide (TMZ). Conclusion This study establishes that miR-423-5p functions as an oncogene in glioma tissues by suppressing ING-4 and suggests that it has therapeutic potential for glioma.