MicroRNA Profile of Exosomes and Parental Cells is Differently Affected by Ionizing Radiation
MicroRNA Profile of Exosomes and Parental Cells is Differently Affected by Ionizing Radiation
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DOI：
10.1667/rade-20-00007
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发表时间：
2020-08-01
影响因子：
3.4
通讯作者：
Widlak, Piotr
中科院分区：
文献类型：
--
作者：
Abramowicz, Agata;Labaj, Wojciech;Widlak, Piotr
外泌体是细胞间通讯的关键介质，参与电离辐射反应的不同方面。外泌体的功能作用取决于它们的分子货物，包括蛋白质和 miRNA 含量。在这项工作中，我们比较了暴露于高剂量辐射的细胞的 miRNA 谱和这些细胞释放的外泌体。 FaDu 细胞（源自人头颈癌）暴露于 2 Gy 和 8 Gy 剂量下，在辐射后 36 小时使用差速离心、超滤和沉淀的组合从培养基中纯化外泌体，然后使用 RNA-seq 方法分析 microRNA。定量了 439 种 miRNA，并且在细胞和外泌体之间观察到它们的相对丰度存在显着差异；外泌体中一些低丰度 miRNA 的表达量过多，而高丰度 miRNA 的表达量不足。在受辐射的细胞和这些细胞释放的外泌体中，有一些 miRNA 种类受到显着影响。然而，在两个 miRNA 组中观察到明显不同的辐射诱导效应，这可以通过 miR-3168 在细胞中显着下调和在外泌体中上调来例证。另一方面，2 Gy 和 8 Gy 的辐射剂量都会产生类似的效果。外泌体中存在的受辐射影响的 miRNA 种类与参与 DNA 损伤和细胞因子介导的反应的基因有关，这可能表明它们在其他地方报道的外泌体介导的辐射诱导旁观者效应中的假设作用。 (C) 2020 年，辐射研究会。
Exosomes are key mediators of cell-to-cell communication involved in different aspects of the response to ionizing radiation. The functional role of exosomes depends on their molecular cargo, including protein and miRNA content. In this work, we compared the miRNA profile of cells exposed to a high-dose of radiation and the exosomes released by those cells. FaDu cells (derived from human head and neck cancer) were exposed to 2 and 8 Gy doses, exosomes were purified from culture media at 36 h postirradiation using a combination of differential centrifugation, ultrafiltration and precipitation, then microRNA was analyzed using the RNA-seq approach. There were 439 miRNA species quantified, and significant differences in their relative abundance were observed between the cells and exosomes; several low-abundance miRNAs were over-represented while high-abundance miRNAwere under-represented in exosomes. There were a few miRNA species markedly affected in irradiated cells and in exosomes released by these cells. However, markedly different radiation-induced effects were observed in both miRNA sets, which could be exemplified by miR-3168 significantly downregulated in cells and upregulated in exosomes. On the other hand, both 2 and 8 Gy radiation doses induced similar effects. Radiation-affected miRNA species present in exosomes are linked to genes involved in the DNA damage and cytokine-mediated response, which may suggest their hypothetical role in the exosome-mediated radiation-induced bystander effect reported elsewhere. (C) 2020 by Radiation Research Society.