Department of Dermatology, Ghent University Hospital, Ghent, Belgium.
J Invest Dermatol. 2013 Jan;133(1):201-9. doi: 10.1038/jid.2012.266. Epub 2012 Aug 16.
The current treatments for hyperpigmentation are often associated with a lack of efficacy and adverse side effects. We hypothesized that microRNA (miRNA)-based treatments may offer an attractive alternative by specifically targeting key genes in melanogenesis. The aim of this study was to identify miRNAs interfering with the pigmentary process and to assess their functional role. miRNA profiling was performed on mouse melanocytes after three consecutive treatments involving forskolin and solar-simulated UV (ssUV) irradiation. Sixteen miRNAs were identified as differentially expressed in treated melan-a cells versus untreated cells. Remarkably, a 15-fold downregulation of miR-145 was detected. Overexpression or downregulation of miR-145 in melan-a cells revealed reduced or increased expression of Sox9, Mitf, Tyr, Trp1, Myo5a, Rab27a, and Fscn1, respectively. Moreover, a luciferase reporter assay demonstrated direct targeting of Myo5a by miR-145 in mouse and human melanocytes. Immunofluorescence tagging of melanosomes in miR-145-transfected human melanocytes displayed perinuclear accumulation of melanosomes with additional hypopigmentation of harvested cell pellets. In conclusion, this study has established an miRNA signature associated with forskolin and ssUV treatment. The significant down- or upregulation of major pigmentation genes, after modulating miR-145 expression, suggests a key role for miR-145 in regulating melanogenesis.
目前针对色素沉着过度的治疗方法往往疗效不佳且存在不良反应。我们假设基于 microRNA (miRNA) 的治疗方法可能是一种有吸引力的替代方法,因为它可以特异性地针对黑素生成中的关键基因。本研究的目的是鉴定干扰色素沉着过程的 miRNAs,并评估其功能作用。在连续三次用 forskolin 和模拟太阳光的紫外线 (ssUV) 照射处理后,对小鼠黑素细胞进行 miRNA 谱分析。在经处理的黑素细胞中鉴定出 16 种差异表达的 miRNA。值得注意的是,miR-145 的表达下调了 15 倍。miR-145 在黑素细胞中的过表达或下调分别导致 Sox9、Mitf、Tyr、Trp1、Myo5a、Rab27a 和 Fscn1 的表达减少或增加。此外,荧光素酶报告基因测定显示 miR-145 可在小鼠和人黑素细胞中直接靶向 Myo5a。转染 miR-145 的人黑素细胞中 melanosomes 的免疫荧光标记显示 melanosomes 在核周聚集,并且收获的细胞沉淀中出现额外的色素减退。总之,本研究建立了与 forskolin 和 ssUV 处理相关的 miRNA 特征。miR-145 表达调节后主要色素沉着基因的下调或上调表明 miR-145 在调节黑素生成中起关键作用。