Department of Plastic Surgery, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.
PLoS One. 2020 Aug 12;15(8):e0237023. doi: 10.1371/journal.pone.0237023. eCollection 2020.
Melanocytes play a central role in skin homeostasis. In this study, we focus on the function of melanocyte releasing exosomes as well as exosomal microRNAs (miRNAs) and investigate whether ultraviolet B (UVB) irradiation exerts an impact on it.
Exosomes derived from human primary melanocytes were isolated through differential centrifugation and were identified in three ways, including transmission electron microscopy observation, nanoparticle tracking analysis, and Western blot analysis. Melanocytes were irradiated with UVB for the indicated time, and then melanin production and exosome secretion were measured. The exosomal miRNA expression profile of melanocytes were obtained by miRNA sequencing and confirmed by real-time PCR.
Exosomes derived from human primary melanocytes were verified. UVB irradiation induced melanin production and increased the exosome release by the melanocytes. In total, 15 miRNAs showed higher levels in UVB-irradiated melanocyte-derived exosomes compared with non-irradiated ones, and the top three upregulated exosomal miRNAs were miR-4488, miR-320d, and miR-7704 (fold change > 4.0).
It is verified for the first time that UVB irradiation enhanced the secretion of exosomes by melanocytes and changed their exosomal miRNA profile. This findings open a new direction for investigating the communication between melanocytes and other skin cells, and the connection between UVB and skin malignant initiation.
黑素细胞在皮肤稳态中起核心作用。在本研究中,我们专注于黑素细胞释放的外泌体的功能以及外泌体 microRNAs(miRNAs),并研究紫外线 B(UVB)照射是否对此产生影响。
通过差速离心分离人原代黑素细胞来源的外泌体,并通过透射电子显微镜观察、纳米颗粒跟踪分析和 Western blot 分析三种方法进行鉴定。用 UVB 照射黑素细胞指定时间,然后测量黑素生成和外泌体分泌。通过 miRNA 测序获得黑素细胞外泌体的 miRNA 表达谱,并通过实时 PCR 进行验证。
验证了来源于人原代黑素细胞的外泌体。UVB 照射诱导黑素生成,并增加黑素细胞的外泌体释放。总共 15 种 miRNA 在 UVB 照射的黑素细胞来源的外泌体中表达水平高于未照射的外泌体,上调最明显的三种外泌体 miRNA 是 miR-4488、miR-320d 和 miR-7704(倍数变化>4.0)。
首次证实 UVB 照射增强了黑素细胞外泌体的分泌,并改变了它们的外泌体 miRNA 谱。这一发现为研究黑素细胞与其他皮肤细胞之间的通讯以及 UVB 与皮肤恶性起始之间的联系开辟了新的方向。