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首页> 外文期刊>Journal of dermatological science >Identification of transcriptional targets of Wnt/beta-catenin signaling in dermal papilla cells of human scalp hair follicles: EP2 is a novel transcriptional target of Wnt3a.
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Identification of transcriptional targets of Wnt/beta-catenin signaling in dermal papilla cells of human scalp hair follicles: EP2 is a novel transcriptional target of Wnt3a.

机译:鉴定人头皮毛囊的真皮乳头细胞中Wnt /β-catenin信号转导的转录靶标:EP2是Wnt3a的新型转录靶标。

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BACKGROUND: Recent studies showed that Wnt signaling through the beta-catenin pathway (canonical Wnt signaling) act on mouse dermal papilla cells (DPCs) enabling hair follicles to keep growing. OBJECTIVE: To investigate whether human DPCs respond to canonical Wnt signaling and, if so, to identify target genes of Wnt/beta-catenin pathway. METHODS: Cultured human DPCs were transiently transfected with the beta-catenin responsive TCF reporter plasmid (pTopflash) and corresponding negative control reporter (pFopflash) to assess the activity of beta-catenin signaling by Wnt3a (one of the canonical Wnts). Immunofluorescence staining was also performed to localize beta-catenin in the presence or absence of Wnt3a. Microarray was carried out using Affymetrix gene chips. RT-PCR analysis and immunoblot were employed to verify microarray data. Cyclic AMP (cAMP) levels were measured using EIA assay after Wnt3a and PGE2 treatment in DPCs. RESULTS: Wnt3a significantly stimulated the transcriptional activity of pTopflash but not pFopflash. In line with this, we identified a number of genes that are regulated by Wnt3a. Some of the differently expressed genes including EP2 were confirmed by RT-PCR analysis. Immunoblot further confirmed that EP2 protein is indeed increased by Wnt3a. DPCs pretreated with Wnt3a showed higher responsiveness to PGE2 as measured by cAMP levels. CONCLUSIONS: Elucidation of the role of Wnt3a-regulated genes identified in this study including EP2 would help our understanding of hair-induction and maintenance of anagen phase.
机译:背景:最近的研究表明,通过β-catenin途径的Wnt信号(经典Wnt信号)作用于小鼠真皮乳头细胞(DPC),从而使毛囊保持生长。目的:研究人类DPCs是否对经典的Wnt信号作出反应,如果是,则确定Wnt /β-catenin途径的靶基因。方法:用β-catenin反应性TCF报告质粒(pTopflash)和相应的阴性对照报告基因(pFopflash)瞬时转染培养的人DPC,以评估Wnt3a(一种典型的Wnt)对β-catenin信号传导的活性。在存在或不存在Wnt3a的情况下,还进行了免疫荧光染色以定位β-catenin。使用Affymetrix基因芯片进行微阵列。 RT-PCR分析和免疫印迹用于验证微阵列数据。在DPC中进行Wnt3a和PGE2处理后,使用EIA分析测量了环AMP(cAMP)水平。结果:Wnt3a显着刺激pTopflash的转录活性,但不刺激pFopflash。与此相符,我们鉴定了许多受Wnt3a调控的基因。通过RT-PCR分析证实了包括EP2在内的一些不同表达的基因。免疫印迹进一步证实,Wnt3a确实增加了EP2蛋白。用cnt水平测量,用Wnt3a预处理的DPC对PGE2的响应性更高。结论:阐明本研究中鉴定的Wnt3a调控基因(包括EP2)的作用将有助于我们了解毛发生长和生长期的维持。

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