MicroRNA sequencing and molecular mechanisms analysis of the effects of gold nanoparticles on human dermal fibroblasts
MicroRNA sequencing and molecular mechanisms analysis of the effects of gold nanoparticles on human dermal fibroblasts
复制标题
纳米金对人真皮成纤维细胞影响的MicroRNA测序及分子机制分析
DOI：
10.1016/j.biomaterials.2014.10.042
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发表时间：
2015-01-01
期刊：
影响因子：
14
通讯作者：
Wu, Si
中科院分区：
文献类型：
--
作者：
Huang, Yan;Lu, Xiaoying;Wu, Si
本研究旨在从microRNA水平探讨金纳米颗粒（gold nanoparticles，GNP）对人皮肤成纤维细胞（human dermal fibroblasts，HDFs）的作用机制。首先，合成20-nm GNP并测定其对HDF增殖的作用。然后利用SOLiD测序技术获得GNP处理后的microRNA表达谱。将microRNA表达数据与先前获得的mRNA和蛋白质表达数据进行比较，以鉴定microRNA靶mRNA/蛋白质。此外，进行了生物信息学分析和验证实验。最后，在microRNA水平上比较GNP和银纳米颗粒（SNP）对HDF的作用。结果表明，GNP没有细胞毒性，因为在用200 μ mGNP处理1、4和8 h后，202个microRNA差异表达。生物信息学分析表明，这些异常表达的miRNA主要参与代谢过程，参与71条生物学通路，其中包括mRNA加工通路和MAPK信号通路两条关键通路，这两条通路是差异表达的miRNA、靶mRNA和蛋白质同时连接的通路。细胞生物学实验证实，GNP影响能量代谢，但不诱导凋亡，破坏细胞骨架或诱导活性氧（ROS）的产生。在microRNA水平上比较GNP和SNPs对HDFs的作用机制，发现GNP与SNPs不同，影响细胞周期，减弱ATP合成抑制和细胞骨架损伤，抑制细胞凋亡，不导致细胞毒性。这两种纳米颗粒产生活性氧的差异可能部分解释了GNP对HDF没有表现出细胞毒性作用的事实，这与SNP不同。(C)2014爱思唯尔有限公司版权所有。
The aim of this study is to investigate the mechanism of the effects of gold nanoparticles (GNPs) on human dermal fibroblasts (HDFs) at the microRNA level. First, 20-nm GNPs were synthesized and their effect on HDF proliferation was assayed. SOLiD sequencing technology was then utilized to obtain the microRNA expression profile after GNP treatment. The microRNA expression data were compared with previously obtained mRNA and protein expression data to identify the microRNA target mRNAs/proteins. Moreover, bioinformatics analyses and validation experiments were conducted. Lastly, the roles of GNPs and silver nanoparticle (SNPs) on HDFs were compared at the microRNA level. The results showed that GNPs were not cytotoxic as 202 microRNAs were differentially expressed after treatment with 200 mu m GNPs for 1, 4 and 8 h. Bioinformatics analyses revealed that these dysregulated miRNAs mainly functioned in metabolic processes and participated in 71 biological pathways, including two key pathways in which the differentially expressed miRNA, target mRNAs and proteins were simultaneously joined, the mRNA processing pathway and MAPK signaling pathway. Biological experiments in cells confirmed that GNPs affected energy metabolism but did not induce apoptosis, destroy the cytoskeleton or induce reactive oxygen species (ROS) production. Comparing the mechanism of the effects of GNPs and SNPs on HDFs at the microRNA level, it was found that, unlike SNPs, GNPs impacted the cell cycle, weakened the ATP synthesis inhibition and cytoskeleton damage, suppressed apoptosis, and did not lead to cytotoxicity. The difference in ROS production by these two nanoparticles might partially explain the fact that GNPs showed no cytotoxic effects on HDFs, unlike SNPs. (C) 2014 Elsevier Ltd. All rights reserved.