Pongking Thatsanapong, Intuyod Kitti, Thongpon Phonpilas, Thanan Raynoo, Sitthirach Chutima, Chaidee Apisit, Kongsintaweesuk Suppakrit, Klungsaeng Sirinapha, Hongsrichan Nuttanan, Sakonsinsiri Chadamas, Vaeteewoottacharn Kulthida, Kanokmedhakul Somdej, Pinlaor Somchai, Pinlaor Porntip
Biomedical Sciences Program, Graduate School, Khon Kaen University, Khon Kaen 40002, Thailand.
Cholangiocarcinoma Research Institute, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
J Tradit Complement Med. 2024 Apr 17;14(6):622-634. doi: 10.1016/j.jtcme.2024.04.007. eCollection 2024 Nov.
Cholangiocarcinoma (CCA) is usually diagnosed at a late stage, leading to treatment failure. Cannabidiol (CBD), exhibits diverse anti-cancer effects in various cancers, offering avenues for improving CCA treatment. This study investigated the effects of CBD on human CCA cells and the underlying mechanisms and .
The effects of CBD on three CCA cell lines (KKU-213B, KKU-100, KKU-055) were assessed using the SRB assay, clonogenic assay, cell cycle arrest, and 3D holotomography. Morphological changes were examined using transmission electron microscopy, while mitochondrial ROS levels and mitochondrial membrane potential were studied using MitoSOX, JC-1, and DCFH-DA. Cellular senescence induction was evaluated via SA-β-gal staining. Protein associatedwith autophagy and cellular senescence were analyzed using Western blot and/or immunofluorescent assays. A xenograft model demonstrated the anti-tumor activity of CBD and the induction of cellular senescence through immunohistochemistry targeting PCNA, β-gal, and p21.
CBD effectively inhibited CCA cell proliferation, suppressed colony formation and induced G0/G1 phase cell cycle arrest. Morphological examination revealed lipid droplets/vesicles in CCA cell lines. CBD induced autophagy by upregulating LC3BII, downregulating p62, and inhibiting the -PI3K, -AKT, and -mTOR pathways. Additionally, CBD disrupted mitochondrial homeostasis by elevating ROS, reducing membrane potential, and induced cellular senescence by increasing the expression of p53 and p21. results were confirmed by xenograft models. Overall, CBD suppresses proliferation and induces cell death, autophagy and senescence in CCA cells via the PI3K/AKT/mTOR pathway, which indicates a therapeutic option for CCA treatment.
胆管癌（CCA）通常在晚期才被诊断出来，导致治疗失败。大麻二酚（CBD）在多种癌症中表现出多种抗癌作用，为改善CCA治疗提供了途径。本研究调查了CBD对人CCA细胞的影响及其潜在机制。
使用SRB法、克隆形成试验、细胞周期阻滞和3D全息断层扫描评估CBD对三种CCA细胞系（KKU - 213B、KKU - 100、KKU - 055）的影响。使用透射电子显微镜检查形态变化，同时使用MitoSOX、JC - 1和DCFH - DA研究线粒体ROS水平和线粒体膜电位。通过SA-β-半乳糖苷酶染色评估细胞衰老诱导情况。使用蛋白质印迹法和/或免疫荧光测定法分析与自噬和细胞衰老相关的蛋白质。异种移植模型通过针对PCNA、β-半乳糖苷酶和p21的免疫组织化学证明了CBD的抗肿瘤活性和细胞衰老诱导情况。
CBD有效抑制CCA细胞增殖，抑制集落形成并诱导G0/G1期细胞周期阻滞。形态学检查显示CCA细胞系中有脂滴/囊泡。CBD通过上调LC3BII、下调p62以及抑制PI3K、AKT和mTOR途径诱导自噬。此外，CBD通过升高ROS、降低膜电位破坏线粒体稳态，并通过增加p53和p21的表达诱导细胞衰老。异种移植模型证实了这些结果。总体而言，CBD通过PI3K/AKT/mTOR途径抑制CCA细胞增殖并诱导细胞死亡、自噬和衰老，这表明CBD是CCA治疗的一种选择。
