Efficient elimination of pancreatic cancer stem cells by hedgehog/GLI inhibitor GANT61 in combination with mTOR inhibition.
Efficient elimination of pancreatic cancer stem cells by hedgehog/GLI inhibitor GANT61 in combination with mTOR inhibition.
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DOI：
10.1186/s12943-016-0534-2
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发表时间：
2016-06-27
期刊：
影响因子：
37.3
通讯作者：
Takao S
中科院分区：
文献类型：
--
作者：
Miyazaki Y;Matsubara S;Ding Q;Tsukasa K;Yoshimitsu M;Kosai K;Takao S
胰腺癌是最致命的恶性肿瘤之一。需要创新的治疗方法，现在癌症干细胞（CSC）有望成为新疗法的有效靶点。因此，我们研究了刺猬蛋白（Hh）信号在维持胰腺癌细胞CSC样特性中的重要性，以发现控制其独特特性的关键分子。我们的实验使用人胰腺癌细胞系 Capan-1、PANC-1、MIA PaCa-2 和 Capan-1 M9，在含有 10% 胎牛血清的 DMEM/F12 培养基中进行。通过球形成实验、免疫荧光染色、流式细胞术分析和MTT细胞活力实验来研究胰腺癌细胞的分子信号和治疗效果。抑制 Hh 通路可显着降低干细胞标记物 CD133 的表达和球体形成（自我更新能力的指标），证明 CSC 样特性受到抑制。此外，GLI 抑制剂 GANT61 比平滑 (SMO) 抑制剂环杷明更能诱导胰腺癌细胞球体形成和细胞活力的减少。这表明 GLI 转录因子，而不是 SMO 膜蛋白，是 Hh 通路中的关键分子。使用 GANT61 联合抑制 mTOR（胰腺 CSC 中的另一个关键分子），可有效降低细胞活力和抑制剂耐药细胞系的球体形成，显示出对胰腺 CSC 样细胞的强大功效和广泛适用性。因此，这种新型联合治疗可通过靶向胰腺 CSC 来控制胰腺癌。这是首次报道通过双重阻断 Hh/GLI 和 mTOR 信号传导有效消除胰腺癌干细胞样细胞。本文的在线版本 (doi:10.1186/s12943-016-0534-2) 包含补充材料，可供授权用户使用。
Pancreatic cancer is one of the most lethal malignancies. The innovative treatments are required and now the cancer stem cells (CSCs) are expected to be an effective target for novel therapies. Therefore we investigated the significance of hedgehog (Hh) signaling in the maintenance of CSC-like properties of pancreatic cancer cells, in order to discover the key molecules controlling their unique properties. Human pancreatic cancer cell lines, Capan-1, PANC-1, MIA PaCa-2 and Capan-1 M9 were used for our experiments in DMEM/F12 medium containing 10 % fetal bovine serum. Sphere formation assay, immunofluorescence staining, flow cytometric analysis and MTT cell viability assay were performed to investigate molecular signals and the efficacy in the treatment of pancreatic cancer cells. Inhibition of the Hh pathway significantly reduced the expression of stem cell marker CD133 and sphere formation, an index of self-renewal capacity, demonstrating the suppression of CSC-like properties. Moreover, the GLI inhibitor GANT61 induced greater reduction in sphere formation and cell viability of pancreatic cancer cells than the smoothened (SMO) inhibitor cyclopamine. This suggests that GLI transcription factors, but not SMO membrane protein, are the key molecules in the Hh pathway. The treatment using GANT61 in combination with the inhibition of mTOR, which is another key molecule in pancreatic CSCs, resulted in the efficient reduction of cell viability and sphere formation of an inhibitor-resistant cell line, showing the strong efficacy and wide range applicability to pancreatic CSC-like cells. Thus, this novel combination treatment could be useful for the control of pancreatic cancer by targeting pancreatic CSCs. This is the first report of the efficient elimination of pancreatic cancer stem-like cells by the double blockage of Hh/GLI and mTOR signaling. The online version of this article (doi:10.1186/s12943-016-0534-2) contains supplementary material, which is available to authorized users.