Abstract：
Wnt/β-catenin signaling plays important role in cancers. Compound 759 is one of the compounds previously screened to identify inhibitors of the Wnt/β-catenin pathway in A549 cells [Lee et al. in Bioorg Med Chem Lett 20:5900-5904, 2010]. However, the mechanism by which Compound 759 induces the inhibition of the Wnt/β-catenin pathway remains unknown. In our study, we employed various assays to comprehensively evaluate the effects of Compound 759 on lung cancer cells. Our results demonstrated that Compound 759 significantly suppressed cell proliferation and Wnt3a-induced Topflash activity and arrested the cell cycle at the G1 stage. Changes in Wnt/β-catenin signaling-related protein expression, gene activity, and protein stability including Axin, and p21, were achieved through western blot and qRT-PCR analysis. Compound 759 treatment upregulated the mRNA level of p21 and increased Axin protein levels without altering the mRNA expression in A549 cells. Co-treatment of Wnt3a and varying doses of Compound 759 dose-dependently increased the amounts of Axin1 in the cytosol and inhibited β-catenin translocation into the nucleus. Moreover, Compound 759 reduced tumor size and weight in the A549 cell-induced tumor growth in the in vivo tumor xenograft mouse model. Our findings indicate that Compound 759 exhibits potential anti-cancer activity by inhibiting the Wnt/β-catenin signaling pathway through the increase of Axin1 protein stability.
摘要：
Wnt/β-catenin信号在癌症中起重要作用。化合物759是先前筛选用于鉴定A549细胞中Wnt/β-连环蛋白途径的抑制剂的化合物之一[Lee等人。在BioorgMedChemLett20：5900-5904，2010]。然而,化合物759诱导Wnt/β-连环蛋白途径抑制的机制仍然未知。在我们的研究中,我们采用各种试验来综合评价化合物759对肺癌细胞的作用。我们的结果表明,化合物759显著抑制细胞增殖和Wnt3a诱导的Topflash活性,并将细胞周期阻滞在G1期。Wnt/β-catenin信号相关蛋白表达的变化,基因活性，和蛋白质稳定性，包括Axin，通过蛋白质印迹和qRT-PCR分析获得p21。化合物759处理上调p21的mRNA水平并增加Axin蛋白水平而不改变A549细胞中的mRNA表达。Wnt3a和不同剂量的化合物759的共治疗剂量依赖性地增加了细胞质中Axin1的量并抑制β-连环蛋白易位到细胞核中。此外，在体内肿瘤异种移植小鼠模型中,化合物759减少了A549细胞诱导的肿瘤生长中的肿瘤大小和重量。我们的发现表明,化合物759通过增加Axin1蛋白稳定性抑制Wnt/β-连环蛋白信号传导途径而表现出潜在的抗癌活性。
