Wu Xiang-Song, Yang Tian, Gu Jun, Li Mao-Lan, Wu Wen-Guang, Weng Hao, Ding Qian, Mu Jia-Sheng, Bao Run-Fa, Shu Yi-Jun, Cao Yang, Wang Xu-an, Ding Qi-Chen, Dong Ping, Xie Shun-Feng, Liu Ying-Bin
aLaboratory of General Surgery, Department of General Surgery, Xinhua Hospital, Institute of Biliary Tract Disease, affiliated with Shanghai Jiao Tong University School of Medicine, Shanghai, China bDepartment of General Surgery, First Affiliated Hospital, College of Medicine, Shantou University, Shantou, China.
Anticancer Drugs. 2014 Oct;25(9):1007-15. doi: 10.1097/CAD.0000000000000124.
Gallbladder carcinoma is the most common malignancy of the biliary tract and is associated with a very poor outcome. The aim of the present study was to investigate the effects of oxymatrine (OM) on gallbladder cancer cells and the possible mechanism of its effects. The effects of OM on the proliferation of gallbladder cancer cells (GBC-SD and SGC-996) were investigated using cell counting kit-8 and colony formation assays. Annexin V/propidium iodide double staining was performed to investigate whether OM could induce apoptosis in gallbladder cancer cells. The mitochondrial membrane potential (ΔΨm) and expression of apoptosis-associated proteins were evaluated to identify a mechanism for the effects of OM. In addition, the RNA expression of relevant genes was measured by qRT-PCR using the SYBR Green method. Finally, a subcutaneous implantation model was used to verify the effects of OM on tumor growth in vivo. We found that OM inhibited the proliferation of gallbladder cancer cells. In addition, Annexin V/propidium iodide double staining showed that OM induced apoptosis after 48 h and the ΔΨm decreased in a dose-dependent manner after OM treatment. Moreover, the activation of caspase-3 and Bax and downregulation of Bcl-2 and nuclear factor κB were observed in OM-treated cells. Finally, OM potently inhibited in-vivo tumor growth following subcutaneous inoculation of SGC-996 cells in nude mice. In conclusion, OM treatment reduced proliferation and induced apoptosis in gallbladder cancer cells, which suggests that this drug may serve as a novel candidate for adjuvant treatment in patients with gallbladder cancer.
胆囊癌是胆道最常见的恶性肿瘤，预后极差。本研究旨在探讨氧化苦参碱（OM）对胆囊癌细胞的影响及其可能的作用机制。使用细胞计数试剂盒-8和集落形成试验研究OM对胆囊癌细胞（GBC-SD和SGC-996）增殖的影响。进行膜联蛋白V/碘化丙啶双重染色以研究OM是否能诱导胆囊癌细胞凋亡。评估线粒体膜电位（ΔΨm）和凋亡相关蛋白的表达以确定OM作用的机制。此外，使用SYBR Green法通过qRT-PCR测量相关基因的RNA表达。最后，采用皮下植入模型验证OM对体内肿瘤生长的影响。我们发现OM抑制胆囊癌细胞的增殖。此外，膜联蛋白V/碘化丙啶双重染色显示，OM在48小时后诱导凋亡，且OM处理后ΔΨm呈剂量依赖性降低。此外，在OM处理的细胞中观察到caspase-3和Bax的激活以及Bcl-2和核因子κB的下调。最后，在裸鼠皮下接种SGC-996细胞后，OM有效抑制体内肿瘤生长。总之，OM处理可降低胆囊癌细胞的增殖并诱导其凋亡，这表明该药物可能成为胆囊癌患者辅助治疗的新候选药物。
