Salinomycin exerts anti-colorectal cancer activity by targeting the beta-catenin/T-cell factor complex
Salinomycin exerts anti-colorectal cancer activity by targeting the beta-catenin/T-cell factor complex
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Salinomycin 通过靶向 β-连环蛋白/T 细胞因子复合物发挥抗结直肠癌活性
DOI：
10.1111/bph.14770
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发表时间：
2019
影响因子：
7.3
通讯作者：
Lu Desheng
中科院分区：
文献类型：
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作者：
Wang Zhongyuan;Zhou Liang;Xiong Yanpeng;Yu Shubin;Li Huan;Fan Jiaoyang;Li Fan;Su Zijie;Song Jiaxing;Sun Qi;Liu Shan-Shan;Xia Yuqing;Zhao Liang;Li Shiyue;Guo Fang;Huang Peng;Carson Dennis A;Lu Desheng
背景和目的盐霉素是一种众所周知的人类癌症干细胞（CSC）抑制剂。然而，盐霉素靶向结直肠CSC的分子机制知之甚少。在此，我们研究了盐霉素在结直肠癌细胞和三种肿瘤模型中的潜在抗肿瘤机制。实验方法采用SuperTopFlash报告系统分析了盐霉素对Wnt/β-catenin通路的抑制作用。采用真实的时间PCR评价Wnt靶基因的mRNA表达。采用免疫共沉淀和体外GST pull‐down试验检测盐霉素对β-catenin/TCF 4 E相互作用的影响。用BrdU掺入法和软琼脂集落形成试验检测细胞增殖。通过球体形成试验评估细胞的干细胞性。Salinomycin对结直肠癌的抗肿瘤作用通过结直肠CSC异种移植物、APC min/+转基因小鼠和患者来源的结直肠肿瘤异种移植物进行了评估。Key ResultsSalinomycin阻断了结直肠癌细胞中β-catenin/TCF 4 E复合物的形成，并在体外GST pull-down试验中阻断了β-catenin/TCF 4 E复合物的形成，从而降低了Wnt靶基因的表达。盐霉素还抑制β-catenin/LEF 1或β-catenin/TCF 4 E复合物介导的转录活性，并对结直肠癌细胞的球体形成、增殖和锚定非依赖性生长表现出抑制作用。在大肠癌异种移植瘤和APC min/+转基因小鼠中，盐霉素可显著抑制肿瘤生长和CSC相关Wnt靶基因包括LGR 5的表达。结论与意义盐霉素可通过破坏β-catenin/TCF复合物抑制大肠癌的生长，可能是一种有前景的大肠癌治疗药物。
Background and PurposeSalinomycin is a well‐known inhibitor of human cancer stem cells (CSCs). However, the molecular mechanism(s) by which salinomycin targets colorectal CSCs is poorly understood. Here, we have investigated underlying antitumour mechanisms of salinomycin in colorectal cancer cells and three tumour models.Experimental ApproachThe inhibitory effect of salinomycin on the Wnt/β‐catenin pathway was analysed with the SuperTopFlash reporter system. The mRNA expression of Wnt target genes was evaluated with real‐time PCR. Effects of salinomycin on β‐catenin/TCF4E interaction were examined using co‐immunoprecipitation and an in vitro GST pull‐down assay. Cell proliferation was determined by BrdU incorporation and soft agar colony formation assay. The stemness of the cells was assessed by sphere formation assay. Antitumour effects of salinomycin on colorectal cancers was evaluated with colorectal CSC xenografts, APCmin/+transgenic mice, and patient‐derived colorectal tumour xenografts.Key ResultsSalinomycin blocked β‐catenin/TCF4E complex formation in colorectal cancer cells and in an in vitro GST pull‐down assay, thus decreasing expression of Wnt target genes. Salinomycin also suppressed the transcriptional activity mediated by β‐catenin/LEF1 or β‐catenin/TCF4E complex and exhibited an inhibitory effect on the sphere formation, proliferation, and anchorage‐independent growth of colorectal cancer cells. In colorectal tumour xenografts and APCmin/+transgenic mice, administration of salinomycin significantly reduced tumour growth and the expression of CSC‐related Wnt target genes including LGR5.Conclusions and ImplicationsOur study suggested that salinomycin could suppress the growth of colorectal cancer by disrupting the β‐catenin/TCF complex and thus may be a promising agent for colorectal cancer treatment.