An Axis Involving SNAI1, microRNA-128 and SP1 Modulates Glioma Progression
An Axis Involving SNAI1, microRNA-128 and SP1 Modulates Glioma Progression
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涉及 SNAI1、microRNA-128 和 SP1 的轴调节神经胶质瘤进展
DOI：
10.1371/journal.pone.0098651
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发表时间：
2014-06-24
期刊：
影响因子：
3.7
通讯作者：
Liu, Ning
中科院分区：
文献类型：
--
作者：
Dong, Qingsheng;Cai, Ning;Liu, Ning
背景：胶质母细胞瘤是一种极具侵袭性的疾病，需要更有效的治疗选择。蜗牛锌指1，在许多肿瘤中失调，已被报道与胶质瘤有关。然而，SNAI1在胶质瘤中功能的生物学机制需要进一步研究。方法：采用实时荧光定量PCR检测miR-128 （microRNA-128）表达水平，western blot检测U87、U251细胞及人脑组织中miR-128蛋白表达情况。进行细胞周期、CCK-8、transwell和创面愈合测定。采用双荧光素酶报告基因法鉴定SNAI1和miR-128b调控的机制。miR-128靶向SP1的机制也通过荧光素酶报告基因试验进行了检测。采用免疫组织化学和原位杂交染色法定量测定人胶质瘤样品中SNAI1、SP1和miR-128的表达水平。结果：中国胶质瘤基因组图谱（CGGA）数据显示，SNAI1在胶质瘤中表达上调，我们在正常组织和胶质瘤组织中证实了这一发现。shRNA敲除SNAI1可延缓胶质瘤细胞周期，抑制胶质瘤细胞的增殖和侵袭。CGGA数据显示，SNAI1与miR-128的Pearson相关指数呈负相关。SNAI1通过结合miR-128b特异性启动子基元抑制miR-128b的表达，而miR-128通过结合SP1的3'-未翻译区靶向SP1。此外，miR-128反义寡核苷酸的引入减轻了SNAI1 shRNA诱导的细胞周期阻滞、增殖和侵袭抑制。免疫组织化学和原位杂交分析SNAI1、SP1和miR-128揭示了它们在胶质瘤样本中的表达水平和相关性。结论：我们提出SNAI1/miR-128/SP1轴在胶质瘤进展中起着至关重要的作用，可能成为临床相关的治疗靶点。
Background: Glioblastoma is an extraordinarily aggressive disease that requires more effective therapeutic options. Snail zinc finger 1, dysregulated in many neoplasms, has been reported to be involved in gliomas. However, the biological mechanisms underlying SNAI1 function in gliomas need further investigation.Methods: Quantitative real-time PCR was used to measure microRNA-128 (miR-128) expression level and western blot was performed to detect protein expression in U87 and U251 cells and human brain tissues. Cell cycle, CCK-8, transwell and wound-healing assays were performed. Dual-luciferase reporter assay was used for identifying the mechanism of SNAI1 and miR-128b regulation. The mechanism of miR-128 targeting SP1 was also tested by luciferase reporter assay. Immunohistochemistry and in situ hybridisation staining were used for quantifying SNAI1, SP1 and miR-128 expression levels in human glioma samples.Results: The Chinese Glioma Genome Atlas (CGGA) data revealed that SNAI1 was up-regulated in glioma and we confirmed the findings in normal and glioma tissues. SNAI1 depletion by shRNA retarded the cell cycle and suppressed proliferation and invasion in glioma cell lines. The CGGA data showed that the Pearson correlation index between SNAI1 and miR-128 was negatively correlated. SNAI1 suppressed miR-128b expression by binding to the miR-128b specific promoter motif, and miR-128 targeted SP1 via binding to the 3'-untranslated region of SP1. Moreover, introduction of miR-128 anti-sense oligonucleotide alleviated the cell cycle retardation, proliferation and invasion inhibition induced by SNAI1 shRNA. Immunohistochemistry and in situ hybridisation analysis of SNAI1, SP1 and miR-128 unraveled their expression levels and correlations in glioma samples.Conclusions: We propose that the SNAI1/miR-128/SP1 axis, which plays a vital role in glioma progression, may come to be a clinically relevant therapeutic target.