Antitumor activity of natural compounds, curcumin and PKF118-310, as Wnt/β-catenin antagonists against human osteosarcoma cells
Antitumor activity of natural compounds, curcumin and PKF118-310, as Wnt/β-catenin antagonists against human osteosarcoma cells
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DOI：
10.1007/s10637-009-9311-z
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发表时间：
2010-12-01
影响因子：
3.4
通讯作者：
Ee, Pui-Lai Rachel
中科院分区：
文献类型：
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作者：
Leow, Pay-Chin;Tian, Quan;Ee, Pui-Lai Rachel
Wnt/β-catenin信号通路的异常激活促进了骨肉瘤的发生和转移。在这项研究中，我们验证了这样的假设，即骨肉瘤的进展可能通过使用小分子抑制剂如姜黄素和PKF118-310扰乱Wnt/β-catenin途径来延迟。荧光素酶报告实验显示，姜黄素和PKF118-310对骨肉瘤细胞内固有的和激活的β-连环蛋白/Tcf转录活性均有抑制作用，表明姜黄素和PKF118-310有效地抑制了Wnt/β-连环蛋白途径。Western印迹分析显示胞质中的β-连环蛋白的含量没有变化，尽管核内的β-连环蛋白在这两种化合物中都显著减少。接下来，我们进行了伤口愈合和Matrigel侵袭试验，观察到姜黄素和PKF118-310治疗对骨肉瘤细胞的迁移和侵袭有剂量依赖性的减少。野生型β-连环素在骨肉瘤细胞中的过表达导致细胞侵袭力增强，但这一作用可被姜黄素显着克服。明胶酶谱和Western blotting结果显示，姜黄素和PKF118-310处理可降低细胞侵袭力，与Wnt/β-catenin内在或外源性激活条件下基质金属蛋白酶-9的活性和蛋白水平有关。通过细胞凋亡实验和细胞周期分析，进一步证明PKF118-310的抗增殖作用与其诱导细胞凋亡和G2/M期阻滞有关。最后，我们观察到这些抗癌作用与细胞周期蛋白D1、c-Myc和Survivin表达的降低有关。我们的研究结果有力地表明姜黄素和PKF118-310在治疗骨肉瘤方面具有很大的治疗潜力。
Aberrant activation of the Wnt/beta-catenin signaling pathway promotes osteosarcoma tumorigenesis and metastasis. In this study, we tested the hypothesis that osteosarcoma progression may be delayed by disrupting the Wnt/beta-catenin pathway using small molecule inhibitors such as curcumin and PKF118-310. Effective inhibitions of the Wnt/beta-catenin pathway by curcumin and PKF118-310 in osteosarcoma cells were shown by the suppression of both intrinsic and activated beta-catenin/Tcf transcriptional activities using luciferase reporter assays. Western blot analysis revealed that there was no change in the amount of cytosolic beta-catenin, although nuclear beta-catenin was markedly reduced by treatment with either compounds. We next performed wound healing and Matrigel invasion assays and observed a dose-dependent decrease in osteosarcoma cell migration and invasion with curcumin and PKF118-310 treatment. Overexpression of the wild-type beta-catenin plasmid in osteosarcoma cells resulted in enhanced cell invasiveness but this effect was significantly overcome by curcumin. Gelatin zymography and Western blotting showed that reduced cell invasion with curcumin and PKF118-310 treatment correlated with the activity and protein level of matrix metalloproteinase-9 under conditions of intrinsic or extrinsic Wnt/beta-catenin activation. Using cell apoptosis assay and cell cycle analysis, we further showed that the anti-proliferative effect of PKF118-310 is attributed to PKF118-310-induced apoptosis and G2/M phase arrest. Lastly, we observed that these anti-cancer effects correlated with the decreased expression of cyclin D1, c-Myc and survivin. Our findings strongly suggest that curcumin and PKF118-310 have great therapeutic potential for the treatment of osteosarcoma.