Mondal Jesmin, Panigrahi Ashis Kumar, Khuda-Bukhsh Anisur Rahman
Department of Zoology, Cytogenetics and Molecular Biology Laboratory, University of Kalyani, Kalyani, West Bengal, India.
Department of Zoology, Fisheries and Aquaculture Laboratory, University of Kalyani, Kalyani, West Bengal, India.
Pharmacogn Mag. 2014 Aug;10(Suppl 3):S524-33. doi: 10.4103/0973-1296.139792.
Conium maculatum extract is used as a traditional medicine for cervix carcinoma including homeopathy. However, no systematic work has so far been carried out to test its anti-cancer potential against cervix cancer cells in vitro. Thus, in this study, we investigated whether ethanolic extract of conium is capable of inducing cytotoxicity in different normal and cancer cell lines including an elaborate study in HeLa cells.
Conium's effects on cell cycle, reactive oxygen species (ROS) accumulation, mitochondrial membrane potential (MMP) and apoptosis, if any, were analyzed through flow cytometry. Whether Conium could damage DNA and induce morphological changes were also determined microscopically. Expression of different proteins related to cell death and survival was critically studied by western blotting and ELISA methods. If Conium could interact directly with DNA was also determined by circular dichroism (CD) spectroscopy.
Conium treatment reduced cell viability and colony formation at 48 h and inhibited cell proliferation, arresting cell cycle at sub-G stage. Conium treatment lead to increased generation of reactive oxygen species (ROS) at 24 h, increase in MMP depolarization, morphological changes and DNA damage in HeLa cells along with externalization of phosphatidyl serine at 48 hours. While cytochrome c release and caspase-3 activation led HeLa cells toward apoptosis, down-regulation of Akt and NFkB inhibited cellular proliferation, indicating the signaling pathway to be mediated via the mitochondria-mediated caspase-3-dependent pathway. CD-spectroscopy revealed that Conium interacted with DNA molecule.
Overall results validate anti-cancer potential of Conium and provide support for its use in traditional systems of medicine.
毒参提取物被用作包括顺势疗法在内的宫颈癌传统药物。然而，迄今为止尚未开展系统工作来测试其对宫颈癌细胞的体外抗癌潜力。因此，在本研究中，我们调查了毒参乙醇提取物是否能够在不同的正常和癌细胞系中诱导细胞毒性，包括对HeLa细胞进行详尽研究。
通过流式细胞术分析毒参对细胞周期、活性氧（ROS）积累、线粒体膜电位（MMP）和凋亡（如有）的影响。还通过显微镜检查确定毒参是否会损伤DNA并诱导形态变化。通过蛋白质印迹法和酶联免疫吸附测定法严格研究与细胞死亡和存活相关的不同蛋白质的表达。还通过圆二色（CD）光谱法确定毒参是否能直接与DNA相互作用。
毒参处理在48小时时降低细胞活力和集落形成，并抑制细胞增殖，使细胞周期停滞在亚G期。毒参处理导致24小时时活性氧（ROS）生成增加，MMP去极化增加，HeLa细胞出现形态变化和DNA损伤，以及48小时时磷脂酰丝氨酸外化。虽然细胞色素c释放和半胱天冬酶-3激活导致HeLa细胞凋亡，但Akt和NFkB的下调抑制细胞增殖，表明信号通路是通过线粒体介导的半胱天冬酶-3依赖性途径介导的。CD光谱显示毒参与DNA分子相互作用。
总体结果证实了毒参的抗癌潜力，并为其在传统医学体系中的应用提供了支持。
