Wang Chaofan, Du Yimei, Lu Changpei, Bi Lingbo, Ding Yunbu, Fan Weixin
Department of Dermatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210000, Jiangsu, China.
Hospital for Skin Diseases, Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing, 210000, Jiangsu, China.
Regen Ther. 2024 Dec 16;28:161-168. doi: 10.1016/j.reth.2024.12.001. eCollection 2025 Mar.
Secreted frizzled-related protein 1 (SFRP1) inhibits Wnt signaling and is differentially expressed in human hair dermal papilla cells (DPCs). However, the specific effect of SFRP1 on cell function remains unclear. Telomerase reverse transcriptase (TERT) representing telomerase activity was found highly active around the hair dermal papilla. TERT levels can be enhanced by activation of the Wnt pathway in cancer cells and embryonic stem cells. Whether this regulatory mechanism is still present in DPCs has not been studied so far.
In this study, DNA plasmids and siRNAs were constructed against the SFRP1 gene and transfected into DPCs cultured in vitro. We detected the viability, proliferation, and migration of DPCs by Calcein/PI fluorescence, CCK-8, -well, or cell scratch experiments, and the expression of potential target genes was also determined through quantitative detection of RNA and protein.
The results demonstrate a significant difference in SFRP1 levels from the control group, suggesting successful transfection of the DNA plasmid and siRNA of SFRP1 into IDPCs. Also, SFRP1 regulates the cell proliferation capacity of IDPCs and reduces their migration functions. The DPCs' living activity, proliferation, and migration function exhibited a negative correlation with the level of SFRP1. SFPR1 also inhibits the protein or RNA expression of β-catenin and TERT in DPCs.
It was proven that in human DPCs, different levels of SFRP1 change how cells work and control Wnt/β-catenin signaling or telomerase activity. This means that blocking SFRP1 could become a new way to treat hair loss diseases in the future.
分泌型卷曲相关蛋白1(SFRP1)抑制Wnt信号传导,且在人毛发真皮乳头细胞(DPCs)中差异表达。然而,SFRP1对细胞功能的具体作用仍不清楚。代表端粒酶活性的端粒酶逆转录酶(TERT)在毛发真皮乳头周围被发现高度活跃。在癌细胞和胚胎干细胞中,Wnt信号通路的激活可提高TERT水平。到目前为止,这种调节机制在DPCs中是否仍然存在尚未得到研究。
在本研究中,构建了针对SFRP1基因的DNA质粒和小干扰RNA(siRNAs),并将其转染到体外培养的DPCs中。我们通过钙黄绿素/碘化丙啶(Calcein/PI)荧光、CCK-8、96孔板或细胞划痕实验检测DPCs的活力、增殖和迁移情况,并通过RNA和蛋白质的定量检测来确定潜在靶基因的表达。
结果表明,与对照组相比,SFRP1水平存在显著差异,这表明SFRP1的DNA质粒和siRNA已成功转染到IDPCs中。此外,SFRP1调节IDPCs的细胞增殖能力并降低其迁移功能。DPCs的生存活性、增殖和迁移功能与SFRP1水平呈负相关。SFPR1还抑制DPCs中β-连环蛋白和TERT的蛋白质或RNA表达。
已证明,在人DPCs中,不同水平的SFRP1会改变细胞的工作方式,并控制Wnt/β-连环蛋白信号传导或端粒酶活性。这意味着阻断SFRP1可能成为未来治疗脱发疾病的新方法。