Novel Effect of Antihelminthic Niclosamide on S100A4-Mediated Metastatic Progression in Colon Cancer
Novel Effect of Antihelminthic Niclosamide on S100A4-Mediated Metastatic Progression in Colon Cancer
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DOI：
10.1093/jnci/djr190
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发表时间：
2011-07-01
影响因子：
10.3
通讯作者：
Stein, Ulrike
中科院分区：
文献类型：
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作者：
Sack, Ulrike;Walther, Wolfgang;Stein, Ulrike
背景结肠癌的转移形成严重降低了患者的生存率。S100 A4是一种钙结合蛋白，与促进结肠癌的转移形成有关。方法为了鉴定S100 A4的转录抑制剂，使用表达S100 A4启动子驱动的荧光素酶（LUC）报告基因构建体（HCT 116-S1004 p-LUC）的人结肠癌细胞系进行1280种具有生物活性的化合物的高通量筛选。氯硝柳胺是一种驱虫剂，被确定为潜在的候选药物。用1 μ M氯硝柳胺处理结肠癌细胞系（HCT 116、SW 620、LS 174 T、SW 480和DLD-1），通过定量逆转录聚合酶链反应和免疫印迹分析对S100 A4 mRNA和蛋白表达的影响，并在体外评估对细胞迁移、侵袭、增殖和集落形成的影响。通过体内成像在人结肠癌的异种移植小鼠模型（n = 8只小鼠）中评估氯硝柳胺对肝转移的作用。通过评分量化氯硝柳胺对停止治疗后转移形成的长期影响，并通过Kaplan-Meier方法分析停止治疗后的总存活率（n = 12只小鼠）。结果氯硝柳胺处理的结肠癌细胞S100 A4 mRNA和蛋白表达降低，细胞迁移、侵袭、增殖和集落形成受到抑制。氯硝柳胺处理的小鼠的体内成像显示与溶剂处理的对照小鼠相比肝转移减少（每组n = 4只小鼠）。与对照组相比，停药26天显示小鼠肝转移形成减少（每组n = 6只小鼠）（对照组vs间断治疗组，平均转移评分= 100% vs 34.9%，平均差异= 65.1%; 95%置信区间[CI] = 18.4%至111.9%，P
Background Metastasis formation in colon cancer severely reduces the survival rate in patients. S100A4, a calcium-binding protein, is implicated in promoting metastasis formation in colon cancer.Methods To identify a transcription inhibitor of S100A4, high-throughput screening of 1280 pharmacologically active compounds was performed using a human colon cancer cell line expressing a S100A4 promoter-driven luciferase (LUC) reporter gene construct (HCT116-S1004p-LUC). Niclosamide, an antihelminthic agent, was identified as a potential candidate. Colon cancer cell lines (HCT116, SW620, LS174T, SW480, and DLD-1) were treated with 1 mu M niclosamide to analyze the effect on S100A4 mRNA and protein expression by quantitative reverse transcription-polymerase chain reaction and immunoblot assays, and effects on cell migration, invasion, proliferation, and colony formation were also assessed in vitro. The effect of niclosamide on liver metastasis was assessed in a xenograft mouse model of human colon cancer (n = 8 mice) by in vivo imaging. The long-term effect of niclosamide on metastasis formation after discontinued treatment was quantified by scoring, and overall survival (n = 12 mice) was analyzed by Kaplan-Meier method after discontinuation of treatment. All statistical tests were two-sided.Results Reduced S100A4 mRNA and protein expression, and inhibited cell migration, invasion, proliferation, and colony formation were observed in niclosamide-treated colon cancer cells in vitro. In vivo imaging of niclosamide-treated mice showed reduced liver metastasis compared with solvent-treated control mice (n = 4 mice per group). Compared with the control group, discontinuation of treatment for 26 days showed reduced liver metastasis formation in mice (n = 6 mice per group) (control vs discontinuous treatment, mean metastasis score = 100% vs 34.9%, mean difference = 65.1%; 95% confidence interval [CI] = 18.4% to 111.9%, P