Wnt blockers inhibit the proliferation of lung cancer stem cells.
Wnt blockers inhibit the proliferation of lung cancer stem cells.
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DOI：
10.2147/dddt.s76602
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Han B
中科院分区：
文献类型：
--
作者：
Zhang X;Lou Y;Zheng X;Wang H;Sun J;Dong Q;Han B
已有研究证实，Wnt通路激活与非小细胞肺癌复发风险相关。然而，药物阻断Wnt信号通路是否能提供治疗的可能性仍是未知的。本研究的目的是评价Wnt信号通路抑制剂甲酸吡喃(PP)对肺癌干细胞(LCSCs)的治疗作用。采用集落形成和球体培养的方法，从三种肺癌细胞系：PC9、SPC-A1和A549中扩增LCSCs。在免疫荧光确认细胞的干性后，用PP与另外三种Wnt信号抑制剂：盐霉素、ICG-001和水飞蓟宾比较，进行细胞活力测定。克隆形成实验和基因表达分析进一步证实了PP对LC SCs的作用。SPC-A1和PC9细胞在球体培养条件下均能成功分化为LSCs，而A549细胞则未能分化为LSCs。免疫荧光检测显示，LSCs可表达多能干细胞标志物，包括NANOG、OCT4、KLF5和SOX2，以及Wnt信号通路分子β-catenin和myc。PP对SPC-A1、PC9和A549的半数抑制浓度分别为10 nM、0.44 nM和0.21 nM，明显低于盐霉素、ICG-001和水飞蓟宾。肺癌细胞经PP处理后，集落形成明显减少，多能干细胞信号通路下调。WNT信号抑制剂PP可抑制LC SCs的增殖，WNT信号通路有望成为肺腺癌的治疗或干预靶点。
Previous study has confirmed that the occurrence of Wnt pathway activation is associated with risk of non-small-cell lung cancer recurrence. However, whether the pharmacologic blocking of the Wnt signaling pathway could provide therapeutic possibility remains unknown. The aim of the present study was to evaluate the therapeutic functions of the Wnt signaling pathway inhibitor pyrvinium pamoate (PP) on lung cancer stem cells (LCSCs) in vitro. Colony formation and sphere culture were performed to enrich LCSCs from three lung cancer cell lines: PC9, SPC-A1, and A549. After confirming stemness by immunofluorescence, PP was employed for cell viability assay by comparison with three other kinds of Wnt signaling inhibitor: salinomycin, ICG-001, and silibinin. The effect of PP on LCSCs was further verified by colony formation assay and gene expression analysis. LCSCs were successfully generated by sphere culture from SPC-A1 and PC9 cells, but not A549 cells. Immunofluorescence assay showed that LCSCs could express pluripotent stem cell markers, including NANOG, Oct4, KLF5, and SOX2, and Wnt signaling pathway molecules β-catenin and MYC. Half-maximal inhibitory concentrations of PP on SPC-A1, PC9, and A549 were 10 nM, 0.44 nM, and 0.21 nM, respectively, which are much lower than those of salinomycin, ICG-001, and silibinin. Moreover, significantly decreased colony formation and downregulation of pluripotent stem cell signaling pathway were observed in lung cancer cells after treatment with PP. Wnt signaling inhibitor PP can inhibit proliferation of LCSCs, and the Wnt signaling pathway could be considered a promising therapeutic or interventional target in lung adenocarcinoma.