miRNA microarray profiling in patients with androgenic alopecia and the effects of miR-133b on hair growth
miRNA microarray profiling in patients with androgenic alopecia and the effects of miR-133b on hair growth
复制标题
雄激素性脱发患者的 miRNA 微阵列分析以及 miR-133b 对毛发生长的影响。
DOI：
10.1016/j.yexmp.2020.104589
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发表时间：
2021-02-01
影响因子：
3.6
通讯作者：
Wan, Miaojian
中科院分区：
文献类型：
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作者：
Deng, Wenjia;Hu, Ting;Wan, Miaojian
目的：雄激素性脱发（阿加）是一种常见的脱发，常伴有多种miRNA的异常表达。本研究旨在探讨阿加患者中异常表达的miRNAs及其特异性分子机制。方法：采用miRNAs芯片分析和qRT-PCR验证技术，对阿加患者中异常表达的miRNAs进行筛选和验证。用不同浓度的二氢睾酮（10（-5）、10（-6）、10（-7）和10（-8）mol/L）处理人毛囊10天。采用免疫荧光染色和qRT-PCR研究了DHT对HF生长、增殖和培养的HF中miRNA表达的影响。此外，用Wnt/β-连环蛋白途径活化剂和/或miR-133 b模拟物处理/转染人毛乳头细胞（HDPC），然后使用CCK-8测定来评估HDPC增殖。采用qRT-PCR和Western blotting方法检测Versican、ALP和β-catenin的表达。结果：miRNA芯片分析发现43个miRNA在阿加患者中差异表达，qRT-PCR证实8个miRNA在AGA患者中差异表达。miR 133 b在阿加患者中的表达异常增高。DHT（10 - 5 mol/L）抑制人HF细胞生长，上调miR-133 b表达; DHT（10 - 7 mol/L）诱导人HF细胞生长，下调miR-133 b表达。与对照组相比，miR 133 b mimic转染组HDPC增殖受到抑制，β-catenin表达下调（P <0. 05）。Wnt/beta-catenin信号通路激活剂处理可显著促进HDPC增殖，并上调beta-catenin的表达（P < 0.05）。此外，HDPC的增殖在Wnt/β-catenin通路激活剂和miR-133 b模拟物共同处理的组与对照组之间没有显著差异联合用药组Versican和ALP表达明显低于对照组（P > 0.05（P < 0.05）我们的数据表明雄激素性脱发患者具有特异性的miRNA表达谱，133 b可能抑制Wnt/β-连环蛋白通路并最终调节毛发生长。
Objective: Androgenetic alopecia (AGA), a common alopecia, is often accompanied by abnormal expression of multiple miRNAs. This study aims to investigate abnormally expressed miRNAs in patients with AGA and their specific molecular mechanism.Methods: miRNA microarray profiling and qRT-PCR validation were used to screen and verify abnormally expressed miRNAs in patients with AGA. Human hair follicles (HFs) were treated with different concentrations of dihydmtestosterone (DHT, 10(-5), 10(-6), 10(-7) and 10(-8) mol/L) for 10 days. The effects of DHT on HF growth, proliferation, and miRNA expression in cultured HFs were investigated using immunofluorescence staining and qRT-PCR. Moreover, human dermal papilla cells (HDPCs) were treated/transfected with a Wnt/beta-catenin pathway activator and/or miR-133b mimic, and then the CCK-8 assay was used to evaluate HDPC proliferation. qRT-PCR and Western blotting were used to measure the expression of Versican, ALP and beta-cateninResults: miRNA microarray profiling identified 43 miRNAs that were significantly differentially expressed in AGA patients, and qRT-PCR verified that 8 miRNAs were significantly differentially expressed. The expression of miR133b was abnormally high in AGA patients. DHT (10 -5 mol/L) inhibited human HF growth and upregulated miR-133b expression, and DHT (10 -7 mol/L) induced human HF growth and downregulated miR-133b expression. HDPC proliferation was inhibited, and the expression of beta-catenin was downregulated in the miR133b mimic-transfected group compared with the control group (P < 0.05). Wnt/beta-catenin pathway activator treatment significantly promoted HDPC proliferation and upregulated the expression of beta-catenin (P < 0.05). In addition, the proliferation of HDPCs was not significantly different between the group cotreated with a Wnt/beta-catenin pathway activator and miR-133b mimic, and the control group (P > 0.05), but the expression of Versican and ALP was suppressed in the cotreatment group (P < 0.05)Conclusion: Our data indicated that patients with androgenic alopecia have specific miRNA expression profiles and that the abnormal expression of miR-133b may inactivate the Wnt/beta-catenin pathway and ultimately regulate hair growth.