The majority of microRNAs detectable in serum and saliva is concentrated in exosomes.
The majority of microRNAs detectable in serum and saliva is concentrated in exosomes.
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DOI：
10.1371/journal.pone.0030679
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Illei GG
中科院分区：
文献类型：
--
作者：
Gallo A;Tandon M;Alevizos I;Illei GG
在自身免疫性疾病中使用microRNA（miRNA）作为生物标志物的兴趣越来越大。它们在许多体液中很容易获得，但如果它们自由循环或封装在微泡中，特别是外泌体中，则存在争议。我们研究了血清和唾液中的大多数miRNA是自由循环的还是集中在外来体中。通过超离心从新鲜和冷冻的人血清和唾液中分离外来体。通过定量RT-PCR比较从外泌体沉淀和外泌体耗尽的血清和唾液提取的所选miRNA的量。一些测试的miRNA是普遍表达的，其他的先前被报道为生物标志物。我们包括先前报道的自由循环的miRNA和一些被认为是外来体特异性的miRNA。通过电子显微镜和western blot证实外泌体组分的纯度。与外泌体耗尽的上清液相比，外泌体沉淀中的miRNA浓度始终较高。我们使用等体积或等量的总RNA作为RT-qPCR的输入获得了相同的结果。完整的未分级血清中的miRNA浓度介于外泌体沉淀和外泌体耗尽的上清液之间。在外来体中可检测到的所选miRNA在全血清和外来体耗尽的上清液中不可检测到。外泌体分离提高了从人类生物流体扩增miRNA的灵敏度。外泌体miRNA应该是早期生物标志物研究的起点，以减少涉及低丰度miRNA的假阴性结果的概率，这些结果可能会被未分级的血清或唾液遗漏。
There is an increasing interest in using microRNAs (miRNA) as biomarkers in autoimmune diseases. They are easily accessible in many body fluids but it is controversial if they are circulating freely or are encapsulated in microvesicles, particularly exosomes. We investigated if the majority of miRNas in serum and saliva are free-circulating or concentrated in exosomes. Exosomes were isolated by ultracentrifugation from fresh and frozen human serum and saliva. The amount of selected miRNAs extracted from the exosomal pellet and the exosome-depleted serum and saliva was compared by quantitative RT-PCR. Some miRNAs tested are ubiquitously expressed, others were previously reported as biomarkers. We included miRNAs previously reported to be free circulating and some thought to be exosome specific. The purity of exosome fraction was confirmed by electronmicroscopy and western blot. The concentration of miRNAs was consistently higher in the exosome pellet compared to the exosome-depleted supernatant. We obtained the same results using an equal volume or equal amount of total RNA as input of the RT-qPCR. The concentration of miRNA in whole, unfractionated serum, was between the exosomal pellet and the exosome-depleted supernatant. Selected miRNAs, which were detectable in exosomes, were undetectable in whole serum and the exosome-depleted supernantant. Exosome isolation improves the sensitivity of miRNA amplification from human biologic fluids. Exosomal miRNA should be the starting point for early biomarker studies to reduce the probability of false negative results involving low abundance miRNAs that may be missed by using unfractionated serum or saliva.
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DOI：
10.1038/nrrheum.2010.81
发表时间：
2010-07
期刊：
Nature reviews. Rheumatology
影响因子：
--
作者：
通讯作者：
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影响因子：
2.6
作者：
Tzimagiorgis, Georgios;Michailidou, Evangelia Z.;Kouidou, Sofia
通讯作者：
Kouidou, Sofia
DOI：
10.1073/pnas.0804549105
发表时间：
2008-07-29
影响因子：
11.1
作者：
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通讯作者：
Tewari, Muneesh
DOI：
10.1073/pnas.1019055108
发表时间：
2011-03-22
影响因子：
11.1
作者：
Arroyo, Jason D.;Chevillet, John R.;Tewari, Muneesh
通讯作者：
Tewari, Muneesh
影响因子：
3.5
作者：
Lachenal, Gaelle;Pernet-Gallay, Karin;Sadoul, Remy
通讯作者：
Sadoul, Remy