Selective enrichment of microRNAs in extracellular matrix vesicles produced by growth plate chondrocytes.
Selective enrichment of microRNAs in extracellular matrix vesicles produced by growth plate chondrocytes.
复制标题
在生长板软骨细胞产生的细胞外基质囊泡中的选择性富集。
DOI：
10.1016/j.bone.2016.03.018
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发表时间：
2016-07
期刊：
影响因子：
4.1
通讯作者：
Schwartz Z
中科院分区：
文献类型：
--
作者：
Lin Z;Rodriguez NE;Zhao J;Ramey AN;Hyzy SL;Boyan BD;Schwartz Z
基质囊泡（Matrix vesicles，MV）是钙化细胞外基质中的一种膜细胞器，含有基质加工酶，通过这些酶的作用调节细胞外环境。目前尚不清楚MV是否也是通过RNA物质，特别是微小RNA（miRNA）转移的细胞-细胞通讯的外泌体介体。我们研究了从肋软骨生长区软骨细胞培养物中分离的MV中RNA的存在。结果表明，MV RNA的平均产量为1.93 ± 0.78ng RNA/104细胞，约占亲本细胞总RNA的0.1%。MV RNA被脂质膜很好地保护免受RNA酶的侵害，并且与细胞相比高度富集小RNA分子。此外，MV中编码和非编码小RNA的比例与亲本细胞不同。在我们使用的所有三种miRNA检测平台中一致地观察到特异性miRNA的富集，这表明miRNA被选择性地包装到MV中。MV富集的miRNAs与骨形成相关的不同信号通路有关。这项研究表明，在软骨和骨发育的细胞-细胞通讯中，通过转移特定的miRNA，MV作为“基质体”发挥着重要作用。
Matrix vesicles (MVs) are membrane organelles found in the extracellular matrix of calcifying cells, which contain matrix processing enzymes and regulate the extracellular environment via action of these enzymes. It is unknown whether MVs are also exosomic mediators of cell-cell communication via transfer of RNA material, and specifically, microRNA (miRNA). We investigated the presence of RNA in MVs isolated from cultures of costochondral growth zone chondrocytes. Our results showed that the average yield of MV RNA was 1.93 ± 0.78 ng RNA/104 cells, which was approximately 0.1% of the parent cell's total RNA. MV RNA was well-protected from RNase by the lipid membrane and was highly enriched in small RNA molecules compared to cells. Moreover, coding and non-coding small RNAs in MVs were in proportions that differed from parent cells. Enrichment of specific miRNAs was consistently observed in all three miRNA detection platforms that we used, suggesting that miRNAs are selectively packaged into MVs. MV-enriched miRNAs were related to different signaling pathways associated with bone formation. This study suggests a significant role for MVs as “matrisomes” in cell-cell communication in cartilage and bone development via transfer of specific miRNAs.