Pang Ming-Jie, Yang Zhun, Zhang Xing-Lin, Liu Zhao-Fang, Fan Jun, Zhang Hong-Ying
Department of Otolaryngology, Qingdao Municipal Hospital, Qingdao 266011, China.
Department of Oncology, Qingdao Municipal Hospital, Qingdao 266011, China.
Acta Pharmacol Sin. 2016 Dec;37(12):1623-1640. doi: 10.1038/aps.2016.98. Epub 2016 Oct 3.
Physcion is a major bioactive ingredient in the traditional Chinese medicine Radix et Rhizoma Rhei, which has an anthraquinone chemical structure and exhibits a variety of pharmacological activities including laxative, hepatoprotective, anti-inflammatory, anti-microbial and anti-proliferative effects. In this study we investigated the effect of physcion on human nasopharyngeal carcinoma in vitro and in vivo, as well as the mechanisms underlying the anti-tumor action.
The nasopharyngeal carcinoma cell line CNE2 was treated with physcion, and cell viability was detected using MTT and colony formation assays. Flow cytometry was used to assess the cell cycle arrest, mitochondrial membrane potential loss, apoptosis, autophagy and intracellular ROS generation. Apoptotic cell death was also confirmed by a TUNEL assay. The expression of target or marker molecules was determined using Western blotting. The activity of caspase-3, 8, and 9 was detected with an ELISA kit. A xenograft murine model was used to evaluate the in vivo anti-tumor action of physcion, the mice were administered physcion (10, 20 mg·kg·d, ip) for 30 d.
Treatment with physcion (5, 10, and 20 μmol/L) dose-dependently suppressed the cell viability and colony formation in CNE2 cells. Physcion (10 and 20 μmol/L) dose-dependently blocked cell cycle progression at G phase and induced both caspase-dependent apoptosis and autophagy in CNE2 cells. Furthermore, physcion treatment induced excessive ROS generation in CNE2 cells, and subsequently disrupted the miR-27a/ZBTB10 axis, resulting in repression of the transcription factor Sp1 that was involved in physcion-induced apoptosis and autophagy. Moreover, physcion-induced autophagy acted as a pro-apoptotic factor, and possibly contributed to physcion-induced apoptosis. In the xenograft murine model, administration of physcion dose-dependently suppressed the tumor growth without affecting the body weight. Furthermore, the anti-tumor effects of physcion were correlated with downregulation of Sp1 and suppression of miR-27a in the tumor tissues.
Physcion induces apoptosis and autophagy in human nasopharyngeal carcinoma by targeting Sp1, which was mediated by ROS/miR-27a/ZBTB10 signaling. The results suggest that physcion is a promising candidate for the treatment of human nasopharyngeal carcinoma.
大黄素是中药大黄中的主要生物活性成分，具有蒽醌化学结构，表现出多种药理活性，包括通便、保肝、抗炎、抗菌和抗增殖作用。在本研究中，我们研究了大黄素在体外和体内对人鼻咽癌的影响以及抗肿瘤作用的潜在机制。
用大黄素处理人鼻咽癌细胞系CNE2，采用MTT法和集落形成试验检测细胞活力。流式细胞术用于评估细胞周期阻滞、线粒体膜电位丧失、凋亡、自噬和细胞内活性氧生成。TUNEL试验也证实了凋亡细胞死亡。用蛋白质免疫印迹法检测靶分子或标志物分子的表达。用ELISA试剂盒检测caspase-3、8和9的活性。采用异种移植小鼠模型评估大黄素的体内抗肿瘤作用，给小鼠腹腔注射大黄素（10、20mg·kg·d），连续30天。
大黄素（5、10和20μmol/L）处理剂量依赖性地抑制CNE2细胞的活力和集落形成。大黄素（10和20μmol/L）剂量依赖性地阻断细胞周期于G期进展，并诱导CNE2细胞发生caspase依赖性凋亡和自噬。此外，大黄素处理诱导CNE2细胞中过量活性氧生成，随后破坏miR-27a/ZBTB10轴，导致参与大黄素诱导凋亡和自噬的转录因子Sp1表达受抑制。此外，大黄素诱导的自噬作为一种促凋亡因子，可能促进了大黄素诱导的凋亡。在异种移植小鼠模型中，大黄素给药剂量依赖性地抑制肿瘤生长，而不影响体重。此外，大黄素的抗肿瘤作用与肿瘤组织中Sp1的下调和miR-27a的抑制相关。
大黄素通过靶向Sp1诱导人鼻咽癌细胞凋亡和自噬，这是由活性氧/miR-27a/ZBTB10信号介导的。结果表明大黄素是治疗人鼻咽癌的一个有前途的候选药物。
