Study of Cinobufagin as a Promising Anticancer Agent in Uveal Melanoma Through Intrinsic Apoptosis Pathway
Study of Cinobufagin as a Promising Anticancer Agent in Uveal Melanoma Through Intrinsic Apoptosis Pathway
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DOI：
10.3389/fonc.2020.00325
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发表时间：
2020-04
影响因子：
4.7
通讯作者：
Leilei Zhang;Xiaolin Huang;Tao Guo;Huixue Wang;Haiyan Fan;Li Fang
中科院分区：
文献类型：
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作者：
Leilei Zhang;Xiaolin Huang;Tao Guo;Huixue Wang;Haiyan Fan;Li Fang
葡萄膜黑色素瘤是成人最常见的原发性眼内癌。蟾毒素是一种中药，由亚洲蟾蜍蟾蜍分泌，广泛用于肿瘤治疗。在此，我们探索了蟾毒球蛋白潜在的抗肿瘤功能，并研究了其在UM细胞中的生化机制。通过细胞活力、细胞周期和细胞凋亡实验来确定cinobufagin的抗肿瘤潜能。菌落形成实验证实，蟾毒蛋白具有剂量依赖性的抗肿瘤活性。在体内和体外实验中，我们发现cinobufagin可以诱导细胞凋亡，并上调cleaved caspase-3、cleaved poly（adp -核糖）聚合酶（PARP）和cleaved caspase-9的表达。此外，增加cinobufagin浓度处理后，线粒体内在凋亡途径也被激活，表现为细胞凋亡增加，Bad和Bax表达增加，Bcl-2和Bcl-xl表达降低，OCM1细胞线粒体膜电位（MMP）降低。综上所述，本临床前研究结果表明，蟾毒球蛋白在UM细胞中具有剂量依赖性抑制细胞存活和诱导细胞凋亡的作用，这为蟾毒球蛋白的生化机制及其作为未来UM化疗药物的潜力提供了新的见解。
Uveal melanoma (UM) is the most common primary intraocular carcinoma in adults. Cinobufagin, secreted by the Asiatic toad Bufo gargarizans, is a traditional Chinese medicine, widely used in tumor treatment. Here, we explored the potential antitumor function of cinobufagin and investigated its biochemical mechanisms in UM cells. The antitumor potential of cinobufagin was determined via cell viability, cell cycle, and apoptosis assays. Colony formation assays confirmed that cinobufagin exerted potent antitumor activity in a dose-dependent manner. We found that cinobufagin could induce cell apoptosis and upregulate the expression of cleaved caspase-3, cleaved poly(ADP-ribose) polymerase (PARP), and cleaved caspase-9 in vivo and in vitro. In addition, after treatment with increased concentrations of cinobufagin, the intrinsic mitochondrial apoptosis pathway was also activated, which was demonstrated by increased cell apoptosis with increased expression of Bad and Bax, decreased expression of Bcl-2 and Bcl-xl, and reduced mitochondrial membrane potential (MMP) in OCM1 cells. Taken together, the results of this preclinical study suggest that cinobufagin can both inhibit cell survival and induce cell apoptosis in a dose-dependent manner in UM cells, which provides new insights into the biochemical mechanism of cinobufagin and its potential as a future chemotherapeutic agent for UM.