Department of Dermatology, The Third Affiliated Hospital, Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou 510630, China.
Department of Dermatology, Yuebei People's Hospital, No. 133 South Huimin Road, Shaoguan 512026, Guangdong, China.
Exp Mol Pathol. 2021 Feb;118:104589. doi: 10.1016/j.yexmp.2020.104589. Epub 2020 Dec 5.
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.
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 dihydrotestosterone (DHT, 10, 10, 10 and 10 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/β-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 β-catenin RESULTS: 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 miR-133b was abnormally high in AGA patients. DHT (10 mol/L) inhibited human HF growth and upregulated miR-133b expression, and DHT (10 mol/L) induced human HF growth and downregulated miR-133b expression. HDPC proliferation was inhibited, and the expression of β-catenin was downregulated in the miR-133b mimic-transfected group compared with the control group (P < 0.05). Wnt/β-catenin pathway activator treatment significantly promoted HDPC proliferation and upregulated the expression of β-catenin (P < 0.05). In addition, the proliferation of HDPCs was not significantly different between the group cotreated with a Wnt/β-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/β-catenin pathway and ultimately regulate hair growth.
雄激素性脱发（AGA）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 AGA 患者中异常表达的 miRNA 及其特定的分子机制。
采用 miRNA 微阵列分析和 qRT-PCR 验证筛选和验证 AGA 患者中异常表达的 miRNA。用人二氢睾酮（DHT，10、10、10 和 10 mol/L）处理不同浓度的人毛囊（HFs）10 天，研究 DHT 对培养的 HFs 生长、增殖和 miRNA 表达的影响。免疫荧光染色和 qRT-PCR 用于检测 DHT 对 HF 生长、增殖和培养的 HFs 中 miRNA 表达的影响。此外，用 Wnt/β-catenin 通路激活剂和/或 miR-133b 模拟物处理/转染人真皮乳头细胞（HDPCs），然后用 CCK-8 测定法评估 HDPC 增殖。qRT-PCR 和 Western blot 用于测量 Versican、ALP 和 β-catenin 的表达。
miRNA 微阵列分析鉴定出 43 个在 AGA 患者中差异表达显著的 miRNA，qRT-PCR 验证了 8 个 miRNA 差异表达显著。miR-133b 在 AGA 患者中的表达异常升高。DHT（10 mol/L）抑制人 HF 生长并上调 miR-133b 表达，DHT（10 mol/L）诱导人 HF 生长并下调 miR-133b 表达。与对照组相比，miR-133b 模拟物转染组 HDPC 增殖受到抑制，β-catenin 表达下调（P < 0.05）。Wnt/β-catenin 通路激活剂处理显著促进 HDPC 增殖，上调 β-catenin 表达（P < 0.05）。此外，Wnt/β-catenin 通路激活剂和 miR-133b 模拟物共处理组与对照组 HDPC 增殖无显著差异（P > 0.05），但共处理组 Versican 和 ALP 的表达受到抑制（P < 0.05）。
我们的数据表明，雄激素性脱发患者具有特定的 miRNA 表达谱，miR-133b 的异常表达可能使 Wnt/β-catenin 通路失活，最终调节毛发生长。
