MiRNAs Mediate GDNF-Induced Proliferation and Migration of Glioma Cells
MiRNAs Mediate GDNF-Induced Proliferation and Migration of Glioma Cells
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miRNA 介导 GDNF 诱导的胶质瘤细胞增殖和迁移
DOI：
10.1159/000485883
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Gao, Dian-Shuai
中科院分区：
文献类型：
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作者：
Zhang, Bao-Le;Dong, Fu-Lu;Gao, Dian-Shuai
背景与目的：胶质细胞源性神经营养因子（GDNF）是促进胶质瘤侵袭性生长的重要因子。本研究旨在揭示胶质瘤细胞增殖和迁移的独特机制。方法：以U251胶质瘤细胞为实验对象，筛选GDNF刺激细胞增殖和迁移的最佳浓度和作用时间。采用基因芯片技术检测microRNA（miRNA）表达谱，并通过实时荧光定量聚合酶链反应（PCR）进行验证。利用miRWalk预测差异表达的miRNAs的靶基因，并利用GO和京都基因与基因组百科全书（KEGG）途径分析筛选多个miRNAs共同作用的靶基因。使用独创性途径分析（IPA）构建调控miRNA网络。通过实时荧光定量PCR或mRNA芯片检测差异表达的miRNAs的靶基因表达。结果：50 ng/mL GDNF作用24 h可明显促进U251胶质瘤细胞的增殖和迁移（P < 0.05）。GDNF处理后，hsa-miR-194- 5 p、hsa-miR-152- 3 p、hsa-miR-205- 5 p、hsa-miR-629- 5 p、hsa-miR-3609、hsa-miR-183- 5 p和hsa-miR-487 b-3 p等7种miRNAs表达显著上调（P < 0.05）。这些miRNAs主要通过丝裂原活化蛋白激酶（mitogen-activated protein kinase，MAPK）信号通路、粘着斑通路和胶质瘤信号通路参与信号转导、细胞粘附和细胞周期。其中五种miRNAs（hsa-miR-194- 5 p、hsa-miR-152- 3 p、hsa-miR-205- 5 p、hsa-miR-183- 5 p和hsa-miR-487 b-3 p）共调节TP 53和Akt。GDNF处理后，两种或两种以上上调的miRNAs共同靶向的4个基因的mRNA表达水平显著降低（P < 0.05）。结论：GDNF处理U251胶质瘤细胞后，可显著增加7种参与细胞粘附和细胞周期调控的miRNAs的表达。
Background/Aims: Glial cell line-derived neurotrophic factor (GDNF) is an important factor promoting invasive glioma growth. This study was performed to reveal a unique mechanism of glioma cell proliferation and migration. Methods: Human U251 glioma cells were used to screen the optimal GDNF concentration and treatment time to stimulate proliferation and migration. MicroRNA (MiRNA) expression profiles were detected by microarray and confirmed by real-time polymerase chain reaction (PCR). The target genes of differentially expressed miRNAs were predicted by miRWalk, and those targeted by multiple miRNAs were screened with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. A regulatory miRNA network was constructed using ingenuity pathway analysis (IPA). Target gene expression of differentially expressed miRNAs was examined by real-time PCR or mRNA microarray. Results: The results show that 50 ng/mL GDNF for 24 h significantly promotes U251 glioma cell proliferation and migration (P < 0.05). Seven miRNAs (hsa-miR-194-5p, hsa-miR-152-3p, hsa-miR-205-5p, hsa-miR-629-5p, hsa-miR-3609, hsa-miR-183-5p, and hsa-miR-487b-3p) were significantly up-regulated after GDNF treatment (P < 0.05). These miRNAs are primarily involved in signal transduction, cell adhesion and cell cycle through mitogen-activated protein kinase (MAPK) signaling, focal adhesion and glioma signal pathways. Five of these miRNAs (hsa-miR-194-5p, hsa-miR-152-3p, hsa-miR-205-5p, hsa-miR-183-5p, and hsa-miR-487b-3p) co-regulate TP53 and Akt. mRNA expression levels of four genes co-targeted by two or more up-regulated miRNAs were significantly decreased after GDNF treatment (P < 0.05). Conclusion: GDNF treatment of U251 glioma cells significantly increased the expression of seven miRNAs involved in cell adhesion and the cell cycle.