Human Villous Trophoblasts Express and Secrete Placenta-Specific MicroRNAs into Maternal Circulation via Exosomes
Human Villous Trophoblasts Express and Secrete Placenta-Specific MicroRNAs into Maternal Circulation via Exosomes
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DOI：
10.1095/biolreprod.108.075481
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发表时间：
2009-10-01
影响因子：
3.6
通讯作者：
Takizawa, Toshihiro
中科院分区：
文献类型：
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作者：
Luo, Shan-Shun;Ishibashi, Osamu;Takizawa, Toshihiro
在这项研究中，我们利用人类胎盘组织进行了小RNA文库测序，以鉴定胎盘特异性miRNAs。我们还验证了人类绒毛膜绒毛可以通过外泌体在细胞外分泌mirna的假设，这些外泌体进而进入母体循环。通过小RNA文库测序，大多数胎盘特异性miRNA（如MIR517A）与19号染色体上的一个miRNA簇相连。miRNA簇基因在胎盘发育过程中存在差异表达。随后通过实时PCR和原位杂交验证，绒毛滋养细胞表达胎盘特异性mirna。血浆小RNA文库分析显示，孕妇血浆中胎盘特异性mirna含量丰富。通过实时PCR，我们证实胎盘特异性mirna在分娩后迅速从血浆中清除，表明这些mirna进入母体循环。通过培养滋养细胞BeWo，我们证实了mirna确实通过外泌体在细胞外释放。综上所述，我们的研究结果表明，mirna从人类胎盘合体滋养细胞输出到母体循环中，在那里它们可以靶向母体组织。最后，为了研究胎盘特异性mirna的生物学功能，我们对转染MIR517A的BeWo细胞进行了蛋白质组分析。生物信息学分析表明，该miRNA可能参与肿瘤坏死因子介导的信号传导。我们的数据为人类胎盘的miRNA生物学提供了重要的见解。
In this study, we performed small RNA library sequencing using human placental tissues to identify placenta-specific miRNAs. We also tested the hypothesis that human chorionic villi could secrete miRNAs extracellularly via exosomes, which in turn enter into maternal circulation. By small RNA library sequencing, most placenta-specific miRNAs (e. g., MIR517A) were linked to a miRNA cluster on chromosome 19. The miRNA cluster genes were differentially expressed in placental development. Subsequent validation by real-time PCR and in situ hybridization revealed that villous trophoblasts express placenta-specific miRNAs. The analysis of small RNA libraries from the blood plasma showed that the placenta-specific miRNAs are abundant in the plasma of pregnant women. By real-time PCR, we confirmed the rapid clearance of the placenta-specific miRNAs from the plasma after delivery, indicating that such miRNAs enter into maternal circulation. By using the trophoblast cell line BeWo in culture, we demonstrated that miRNAs are indeed extracellularly released via exosomes. Taken together, our findings suggest that miRNAs are exported from the human placental syncytiotrophoblast into maternal circulation, where they could target maternal tissues. Finally, to address the biological functions of placenta-specific miRNAs, we performed a proteome analysis of BeWo cells transfected with MIR517A. Bioinformatic analysis suggests that this miRNA is possibly involved in tumor necrosis factor-mediated signaling. Our data provide important insights into miRNA biology of the human placenta.