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暴露于电离辐射后血浆外泌体样囊泡中蛋白质和微小RNA含量的定量变化。

Quantitative changes in the protein and miRNA cargo of plasma exosome-like vesicles after exposure to ionizing radiation.

作者信息

Yentrapalli Ramesh, Merl-Pham Juliane, Azimzadeh Omid, Mutschelknaus Lisa, Peters Carsten, Hauck Stefanie M, Atkinson Michael J, Tapio Soile, Moertl Simone

机构信息

a Institute of Radiation Biology , Helmholtz Zentrum München, German Research Center for Environmental Health , Neuherberg , Germany.

e Medical Genetics and Functional, Genomics , Aix Marseille University - Inserm UMR_S 910 , Marseille , France.

出版信息

Int J Radiat Biol. 2017 Jun;93(6):569-580. doi: 10.1080/09553002.2017.1294772. Epub 2017 Mar 7.

Abstract

PURPOSE

Multiple cell types secrete exosome-like extracellular vesicles (ELVs) to the extracellular environment. Pathological conditions can produce characteristic changes to the vesicle cargo. We investigated if ionizing radiation is capable of inducing changes in the protein and microRNA (miRNA) cargo of ELVs.

MATERIALS AND METHODS

Whole blood samples from healthy donors were irradiated with 2 Gy gamma rays and then peripheral blood mononuclear cells and plasma were separated from residual blood and co-cultivated for 24 h. The released ELVs were collected by differential ultracentrifugation from irradiated and non-irradiated samples. microRNAs and proteins were quantified by qPCR and label-free proteomics.

RESULTS

Here we report a first characterization of radiation-induced changes in the protein and miRNA cargo of ELVs isolated from plasma. Proteome analysis of ELVs identified 214 proteins, of which nine significantly changed their abundance after irradiation. The radiation-induced down-regulation of afamin and serpine peptidase F1 was confirmed by immunoblotting. miRNA expression profiling identified 58 different exosomal miRNAs, the expression of miR-204-5p, miR-92a-3p and miR-31-5p was significantly increased in ELVs from irradiated samples.

CONCLUSIONS

This study provides evidence that radiation-induced changes occur in the protein and miRNA cargo of plasma ELVs. These data imply a novel systemic communication pathway between irradiated and non-irradiated cells and tissues.

摘要

目的

多种细胞类型会向细胞外环境分泌外泌体样细胞外囊泡(ELV)。病理状况可使囊泡内容物产生特征性变化。我们研究了电离辐射是否能够诱导ELV的蛋白质和微小RNA(miRNA)内容物发生变化。

材料与方法

对健康供体的全血样本进行2 Gy伽马射线照射,然后从剩余血液中分离出外周血单核细胞和血浆,并共同培养24小时。通过差速超速离心从照射和未照射的样本中收集释放的ELV。通过qPCR和无标记蛋白质组学对微小RNA和蛋白质进行定量分析。

结果

在此我们首次报道了从血浆中分离出的ELV的蛋白质和miRNA内容物辐射诱导变化的特征。对ELV的蛋白质组分析鉴定出214种蛋白质,其中9种在照射后丰度发生显著变化。通过免疫印迹法证实了辐射诱导的afamin和丝氨酸蛋白酶F1的下调。miRNA表达谱分析鉴定出58种不同的外泌体miRNA,照射样本的ELV中miR-204-5p、miR-92a-3p和miR-31-5p的表达显著增加。

结论

本研究提供了证据表明血浆ELV的蛋白质和miRNA内容物会发生辐射诱导的变化。这些数据暗示了照射细胞与未照射细胞及组织之间一种新的全身通讯途径。

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