| 王欣愉,王彩霞,刘倩文,等.基于斑马鱼胚胎表型的Wnt/β-catenin信号通路小分子抑制剂的筛选方法[J].中国海洋药物,2022,41(1):51-56. |
| 基于斑马鱼胚胎表型的Wnt/β-catenin信号通路小分子抑制剂的筛选方法 |
| Establishment and evaluation of a method for screening smallmolecular inhibitors on Wnt/β-catenin signaling pathway |
| 投稿时间：2021-03-16 修订日期：2021-04-25 |
| DOI： |
| 中文关键词: Wnt/β-catenin信号通路 小分子抑制剂 左右不对称 斑马鱼 |
| English Keywords:Wnt/β-catenin signaling pathway small molecule inhibitors left-right asymmetry zebrafish |
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| 中文摘要: |
| 目的 利用模式生物斑马鱼,建立1种基于胚胎表型的直观、快速的小分子抑制剂筛选方法,从而高效筛选和评价小分子化合物对体内Wnt/β-catenin信号通路的抑制活性。方法 Wnt/β-catenin信号通路激活剂6-bromoindirubin-3-oxime (BIO) 处理受精后6小时 (hpf)斑马鱼胚胎,检测胚胎48 hpf眼睛发育情况；Wnt/β-catenin信号通路抑制剂处理6 hpf斑马鱼胚胎,检测48 hpf胚胎心脏不对称发育情况；小分子化合物与BIO共同处理6 hpf斑马鱼胚胎,检测斑马鱼眼部表型营救情况。结果 Wnt/β-catenin信号通路激活剂BIO处理斑马鱼胚胎,48 hpf胚胎出现“无眼”表型；Wnt/β-catenin信号通路抑制剂iCRT3、PNU-74654处理斑马鱼胚胎,48 hpf胚胎心脏左右不对称发育出现异常；Wnt/β-catenin信号通路抑制剂XAV-939、PNU-74654分别与BIO共同处理斑马鱼胚胎,48 hpf能够显著营救由BIO激活所导致斑马鱼的眼部缺失表型。已知Wnt/β-catenin信号通路异常激活与诸多癌症密切相关,因此,本文为筛选具有靶向抑癌作用的先导化合物提供了高效、可靠、快速的筛选方法。 |
| English Summary: |
| Objective To establish a phenotypic screening of small molecule inhibitors on Wnt/β-catenin signaling pathway in vivo, we take advantage of zebrafish embryos as model organism to evaluate phenotypic screening. Methods Zebrafish embryos showed disorder with left-right asymmetry when Wnt action was inhibited. In addition, abnormal activation of Wnt led to "eyeless" phenotype. Addition of small molecular compounds to embryos at 6 hpf resulted in cardiac asymmetry at 48 hpf. Addition of 6-bromoindirubin-3-oxime (BIO) to embryos at 6 hpf resulted in eyeless phenotype at 48 hpf. The inhibitory action of compounds on Wnt/β-catenin signaling pathway was evaluated by phenotype analysis. Results Among the three well-characterized inhibitors of Wnt/β-catenin signaling, XAV-939, iCRT3, and PNU-74654, addition of iCRT3 and PNU-74654 leads to abnormal heart left-right asymmetry at 48 hpf and addition of XAV-939 or PNU-74654 with BIO could restore the "eyeless" phenotype of zebrafish at 48 hpf, suggesting that the methods we established are reliable for screening small molecular compounds involved in inhibition of Wnt/β-catenin signaling pathway. This report provides an efficient and reliable screening method for targeting Wnt/β-catenin signaling pathway. |
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