Xia Zhili, Li Minzhen, Hu Meng, Lin Yanyan, Atteh Lawrence Lawer, Fu Wenkang, Gao Long, Bai Mingzhen, Huang Chongfei, Yue Ping, Liu Yu, Meng Wenbo
The First Clinical Medical College, Lanzhou University, Lanzhou, China.
Department of Gynecology and Obstetrics, West China Second University Hospital, Sichuan University, Chengdu, China.
Front Oncol. 2022 Sep 16;12:982961. doi: 10.3389/fonc.2022.982961. eCollection 2022.
Intrahepatic cholangiocarcinoma (ICC) is a malignant tumor that originates from bile duct's epithelial cells and is usually characterized by insidious symptoms and poor prognosis. Cinobufotalin (CB), an active ingredient obtained from the Traditional Chinese Medicine ChanSu, is purported to exhibit a wide range of antitumorigenic activities. However, the mechanism by which it achieves such pharmacological effects remains elusive. Here, we disclosed the mechanism of action by which CB inhibits ICC cells. Initial experiments revealed that the proliferation of RBE and HCCC-9810 cells was significantly inhibited by CB with IC50 values of 0.342 μM and 0.421 μM respectively. CB induced the expression of caspase-3 subsequently leading to the apoptosis of ICC cells. Phosphoproteomics revealed that the phosphorylation of many proteins associated with DNA damage response increased. Kinase-substrate enrichment analysis revealed that ATM was activated after CB treatment, while CDK1 was inactivated. Activated ATM increased p-CHK2-T68 and p-p53-S15, which promoted the expression of FAS, DR4 and DR5 and triggered cell apoptosis. In summary, this work reveals the role of CB in inducing DNA damage and cell apoptosis involved in the activation of the ATM/CHK2/p53 signaling pathway, and indicates that CB may serve as a chemotherapeutic drug candidate for ICC treatment.
肝内胆管癌（ICC）是一种起源于胆管上皮细胞的恶性肿瘤，通常症状隐匿，预后较差。华蟾素（CB）是从中药蟾酥中提取的一种活性成分，据称具有广泛的抗肿瘤活性。然而，其发挥这种药理作用的机制仍不清楚。在此，我们揭示了CB抑制ICC细胞的作用机制。初步实验表明，CB对RBE和HCCC-9810细胞的增殖有显著抑制作用，IC50值分别为0.342 μM和0.421 μM。CB诱导caspase-3表达，随后导致ICC细胞凋亡。磷酸化蛋白质组学显示，许多与DNA损伤反应相关的蛋白质磷酸化增加。激酶-底物富集分析显示，CB处理后ATM被激活，而CDK1失活。激活的ATM增加p-CHK2-T68和p-p53-S15，促进FAS、DR4和DR5的表达并触发细胞凋亡。总之，这项工作揭示了CB在诱导DNA损伤和细胞凋亡中所起的作用，这涉及ATM/CHK2/p53信号通路的激活，并表明CB可能作为ICC治疗的化疗候选药物。
