GOLPH3 Regulates Exosome miRNA Secretion in Glioma Cells
GOLPH3 Regulates Exosome miRNA Secretion in Glioma Cells
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GOLPH3 调节胶质瘤细胞中外泌体 miRNA 的分泌
DOI：
10.1007/s12031-020-01535-6
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发表时间：
2020-03-29
影响因子：
3.1
通讯作者：
Zhou, Xiuping
中科院分区：
文献类型：
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作者：
Hu, Pengfei;Wang, Kai;Zhou, Xiuping
我们的目的是检查高尔基体蛋白GOLPH 3是否可以影响神经胶质瘤细胞源性外泌体的分泌。通过超速离心从U251和U87细胞培养物的上清液中提取外泌体，并通过透射电子显微镜（TEM）、马尔文分析仪和western印迹进行鉴定。通过测量总蛋白水平和多囊泡体（MVB）（外泌体的来源）的数量来检查外泌体的量。通过高通量测序分析外泌体miRNA，然后进行GO和KEGG分析，并通过qRT-PCR验证。GOLPH 3不影响外泌体的总蛋白水平和MVB的数量。然而，我们发现149个差异表达的miRNAs在载体和GOLPH 3过表达组之间的exosomes中，14个miRNAs仅在GOLPH 3过表达细胞中检测到。这些miRNAs预测的靶基因具有结合和催化活性功能，在细胞内吞、RNA转运、甲状腺激素信号转导和肿瘤中的miRNAs途径中富集。GOLPH 3不影响exosomes的数量，但能促进exosomes中miRNA的表达，这可能在GOLPH 3促进胶质瘤发生发展中起一定作用。
We aimed to examine whether golgi protein GOLPH3 could affect the secretion of glioma cell-derived exosomes. The exosomes were extracted by ultra-centrifugation from the supernatant of U251 and U87 cell cultures and identified by transmission electron microscopy (TEM), Malvern analyzer, and western blot. The quantity of exosomes was examined by measuring the total protein levels and the number of multiple vesicle bodies (MVBs), the source of exosomes. The exosome miRNAs were analyzed by high-throughput sequencing followed by GO and KEGG analysis, and validated by qRT-PCR. GOLPH3 could not affect the total protein levels of exosomes and the number of MVBs. However, we found 149 differentially expressed miRNAs in exosomes between vector and GOLPH3 over-expression group, and 14 miRNAs were only examined in GOLPH3 over-expression cells. The predicted target genes of these miRNAs had functions in binding and catalytic activity, which were enriched in the pathways of endocytosis, RNA transportation, thyroid hormone signaling and miRNAs in cancer. GOLPH3 could not affect the quantity of exosomes, but rather contribute to miRNA expression in exosomes, which may play some functions in the promotion effect of GOLPH3 on glioma development.