Laboratory of Veterinary Pharmacology, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
Laboratory of Veterinary Pathology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
Int J Mol Sci. 2018 Apr 6;19(4):1098. doi: 10.3390/ijms19041098.
Colorectal cancer is one of the most common causes of cancer death worldwide. In patients with metastatic colorectal cancer, combination treatment with several anti-cancer drugs is employed and improves overall survival in some patients. Nevertheless, most patients with metastatic disease are not cured owing to the drug resistance. Cancer stem cells are known to regulate resistance to chemotherapy. In the previous study, we established a novel three-dimensional organoid culture model from tumor colorectal tissues of human patients using an air-liquid interface (ALI) method, which contained numerous cancer stem cells and showed resistance to 5-fluorouracil (5-FU) and Irinotecan. Here, we investigate which inhibitor for stem cell-related signal improves the sensitivity for anti-cancer drug treatment in tumor ALI organoids. Treatment with Hedgehog signal inhibitors (AY9944, GANT61) decreases the cell viability of organoids compared with Notch (YO-01027, DAPT) and Wnt (WAV939, Wnt-C59) signal inhibitors. Combination treatment of AY9944 or GANT61 with 5-FU, Irinotecan or Oxaliplatin decreases the cell viability of tumor organoids compared with each anti-cancer drug alone treatment. Treatment with AY9944 or GANT61 inhibits expression of stem cell markers c-Myc, CD44 and Nanog, likely through the decrease of their transcription factor, GLI-1 expression. Combination treatment of AY9944 or GANT61 with 5-FU or Irinotecan also prevents colony formation of colorectal cancer cell lines HCT116 and SW480. These findings suggest that Hedgehog signals mediate anti-cancer drug resistance in colorectal tumor patient-derived ALI organoids and that the inhibitors are useful as a combinational therapeutic strategy against colorectal cancer.
结直肠癌是全球最常见的癌症死亡原因之一。在转移性结直肠癌患者中，采用几种抗癌药物联合治疗可以提高部分患者的总生存率。然而，由于耐药性的存在，大多数转移性疾病患者无法治愈。癌症干细胞被认为可以调节对化疗的耐药性。在之前的研究中，我们使用气液界面（ALI）方法从人类患者的结直肠肿瘤组织中建立了一种新型的三维类器官培养模型，该模型包含大量的癌症干细胞，并对 5-氟尿嘧啶（5-FU）和伊立替康表现出耐药性。在这里，我们研究了哪种用于干细胞相关信号的抑制剂可以提高肿瘤 ALI 类器官对抗癌药物治疗的敏感性。与 Notch（YO-01027、DAPT）和 Wnt（WAV939、Wnt-C59）信号抑制剂相比，Hedgehog 信号抑制剂（AY9944、GANT61）处理降低了类器官的细胞活力。与每种抗癌药物单独处理相比，AY9944 或 GANT61 与 5-FU、伊立替康或奥沙利铂联合治疗降低了肿瘤类器官的细胞活力。AY9944 或 GANT61 处理抑制了干细胞标志物 c-Myc、CD44 和 Nanog 的表达，可能是通过降低其转录因子 GLI-1 的表达。AY9944 或 GANT61 与 5-FU 或伊立替康联合治疗也阻止了结直肠癌细胞系 HCT116 和 SW480 的集落形成。这些发现表明 Hedgehog 信号在结直肠肿瘤患者衍生的 ALI 类器官中介导了抗癌药物耐药性，并且抑制剂可作为针对结直肠癌的联合治疗策略。
