| 莫丽君,宋曼,刘晓丹,关华,周平坤.γ射线照射BEP2D细胞诱导表达的外泌体miRNA的鉴定和功能生物信息学分析[J].中华放射医学与防护杂志,2018,38(7):481-488 |
| γ射线照射BEP2D细胞诱导表达的外泌体miRNA的鉴定和功能生物信息学分析 |
| Identification and bioinformatic function analysis of differentially expressed miRNAs in the exosomes secreted from BEP2D cells irradiated by γ-rays |
| 投稿时间：2018-05-02 |
| DOI：10.3760/cma.j.issn.0254-5098.2018.07.001 |
| 中文关键词: γ射线 外泌体 miRNA 生物信息学 旁效应 |
| 英文关键词:γ-ray irradiation Exosomes miRNA Bioinformatics Bystander effect |
| 基金项目:国家自然科学基金（81530085） |
|
| 摘要点击次数: 2695 |
| 全文下载次数: 1750 |
| 中文摘要: |
| 目的 识别鉴定γ射线照射细胞外泌体中放射诱导表达的miRNA分子，为揭示放射损伤旁效应机制提供新的线索。方法 60Co γ射线照射人支气管上皮细胞BEP2D，超高速离心法分别从2 Gy照射细胞和对照细胞的培养液上清中收集外泌体，用电子显微镜鉴定外泌体，miRNA芯片杂交检测分析外泌体中miRNA表达谱，qRT-PCR方法验证部分miRNA的表达变化，通过TargetScan、miRanda、GO和KEGG等生物信息学技术分析预测靶基因及其相关功能通路。结果BEP2D细胞2 Gy照射后4 h的外泌体中miRNA表达与对照组相比，共鉴定了16个表达水平显著上调的miRNA分子（P<0.05），对其中miR-100-5p、miR-1246、miR-29b-3p、miR-7-5p在外泌体中表达上调通过定量RT-PCR得到进一步的验证。同时，还分析了上述4个miRNA在受照细胞内的表达变化，结果显示除miR-7-5p外其余3个miRNA分子在照射后2 h表达上升，4 h时后均显著降低，而此时外泌体中相应miRNA的表达是显著增加的。生物信息学分析结果表明，这些miRNA可能通过调控细胞粘附、蛋白磷酸化修饰、mTOR信号通路、染色质修饰以及DNA损伤的HR和NHEJ修复等信号通路来参与细胞辐射旁效应过程。结论 鉴定了辐射诱导BEP2D细胞外泌体中差异表达miRNAs分子，这些miRNA的靶基因参与细胞重要的生物学过程和功能信号通路，为揭示辐射诱导旁效应的机制提供了新的线索。 |
| 英文摘要: |
| Objective To identify the differentially expressed miRNAs in the exosomes secreted from γ-ray irradiated cells and provide new clues in disclosing the mechanisms of radiation-induced bystander effects. Methods The human bronchial epithelial cells (BEP2D) were irradiated with 60Co γ-rays, and the exosomes were collected by ultracentrifugation from the culture medium of 2 Gy-irradiated cells and non-irradiated control. The exosomes were identified by an electron microscopy. The miRNA microarray technique was used to analyze the miRNA expression profiles in the exosomes. qRT-PCR was used to verify the miRNAs expression. The functional pathways of miRNAs targeting genes were predicted by informatic analysis using the databases of TargetScan, miRanda, GO and KEGG. Results Sixteen miRNA with significantly increased expression (P<0.05) were identified in the exosomes of BEP2D cells at 4 h post-2 Gy irradiation as compared with the non-irradiated control cells, among which miR-100-5p, miR-1246, miR-29b-3p, and miR-7-5p were further confirmed to be unregulated by qRT-PCR assay (P<0.05). Meanwhile, the expression changes of above-mentioned four miRNAs were also investigated in the irradiated cells. The data indicated the expression was significantly increased at 2 h post-2 Gy irradiation for the miR-100-5p, miR-1246, miR-29b-3p in addition to miR-7-5p. However, all these four miRNAs were downregulated in the cells at 4 h post-irradiation and then gradually recovered. Bioinformatics analysis showed that the targeted genes of these differentially expressed miRNAs might participate in the biological processes and signal pathways of cell adhesion, mTOR signal pathway, chromatin modification, HR and NHEJ pathways of DNA repair and so on. Conclusions Radiation-inducible miRNAs have been identified in the exosomes from the irradiated BEP2D cells. The target genes of these miRNAs play roles in a series of important biological processes and functional pathways, which provides new clues in elucidating the mechanisms of radiation-induced bystander effects. |
| HTML 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|