大多数胆管癌（CCA）患者吉西他滨治疗失败是由于耐药性。具有最小毒性的天然植物次级化合物（例如大麻二酚（CBD））的治疗潜力是吉西他滨耐药性 CCA 的一个有前景的研究方向。我们的目的是在体外和体内研究CBD对吉西他滨耐药CCA（KKU-213BGemR）细胞的影响。使用MTT实验、克隆形成实验和流式细胞术评估体外细胞增殖、集落形成、细胞凋亡和细胞周期阻滞。 。使用 DCFH-DA 荧光探针评估 CBD 对 ROS 产生的影响。通过蛋白质印迹分析研究了 CBD 对 ER 应激相关细胞凋亡的影响机制。还使用吉西他滨耐药的CCA异种移植模型，并通过免疫组织化学分析评估PCNA和CHOP的表达。CBD对KKU-213BGemR细胞的IC50值范围为19.66至21.05 µM。对于非癌性永生化成纤维细胞系，相关值为 18.29 至 19.21 µM。 CBD 在 10 至 30 µM 范围内以剂量依赖性方式抑制 KKU-213BGemR 细胞的集落形成。 30 µM CBD 显着增加早期 (16.37%) (P = 0.0024) 和晚期 (1.8%) (P < 0.0001) 的细胞凋亡，总计 18.17% 细胞凋亡 (P = 0.0017)，部分原因是增加 ROS 产生（P < 0.0001）。 CBD 10 和 20 µM 时的多相细胞周期停滞在 G0/G1 时显着增加（P = 0.004 和 P = 0.017），而 CBD 30 µM 时的 G2/M 时（P = 0.005）。 CBD 治疗导致 ER 应激相关凋亡蛋白表达增加，包括 p-PERK、BiP、ATF4、CHOP、BAX 和细胞色素 c。在异种移植小鼠中，CBD 在 10 和 40 mg/kg·Bw 时显着抑制肿瘤（分别为 P = 0.0007 和 P = 0.0278），这得到了肿瘤组织中 CHOP 增加但 PCNA 表达降低的支持（P < 0.0001)。结果表明，CBD 在体外和体内对吉西他滨耐药的 CCA 表现出有效的抗癌活性，部分是通过 ER 应激介导的机制实现的。这些结果表明，临床探索性使用 CBD 对吉西他滨耐药的 CCA 患者是有必要的。© 2024。作者。

Failure of treatment with gemcitabine in most cholangiocarcinoma (CCA) patients is due to drug resistance. The therapeutic potential of natural plant secondary compounds with minimal toxicity, such as cannabidiol (CBD), is a promising line of investigation in gemcitabine-resistant CCA. We aim to investigate the effects of CBD on gemcitabine-resistant CCA (KKU-213BGemR) cells in vitro and in vivo.In vitro, cell proliferation, colony formation, apoptosis and cell cycle arrest were assessed using MTT assay, clonogenicity assay and flow cytometry. The effect of CBD on ROS production was evaluated using the DCFH-DA fluorescent probe. The mechanism exerted by CBD on ER stress-associated apoptosis was investigated by western blot analysis. A gemcitabine-resistant CCA xenograft model was also used and the expression of PCNA and CHOP were evaluated by immunohistochemical analysis.The IC50 values of CBD for KKU-213BGemR cells ranged from 19.66 to 21.05 µM. For a non-cancerous immortalized fibroblast cell line, relevant values were 18.29 to 19.21 µM. CBD suppressed colony formation by KKU-213BGemR cells in a dose-dependent manner in the range of 10 to 30 µM. CBD at 30 µM significantly increased apoptosis at early (16.37%) (P = 0.0024) and late (1.8%) stages (P < 0.0001), for a total of 18.17% apoptosis (P = 0.0017), in part by increasing ROS production (P < 0.0001). Multiphase cell cycle arrest significantly increased at G0/G1 with CBD 10 and 20 µM (P = 0.004 and P = 0.017), and at G2/M with CBD 30 µM (P = 0.005). CBD treatment resulted in increased expression of ER stress-associated apoptosis proteins, including p-PERK, BiP, ATF4, CHOP, BAX, and cytochrome c. In xenografted mouse, CBD significantly suppressed tumors at 10 and 40 mg/kg·Bw (P = 0.0007 and P = 0.0278, respectively), which was supported by an increase in CHOP, but a decrease in PCNA expression in tumor tissues (P < 0.0001).The results suggest that CBD exhibits potent anti-cancer activity against gemcitabine-resistant CCA in vitro and in vivo, in part via ER stress-mediated mechanisms. These results indicate that clinical explorative use of CBD on gemcitabine-resistant CCA patients is warranted.© 2024. The Author(s).