Tripathi Surya Kant, Biswal Bijesh K
Cancer and Stem Cell Laboratory, Department of Life Science, National Institute of Technology, Rourkela, Sundargarh, Odisha, 769008, India.
Mol Biol Rep. 2018 Dec;45(6):2283-2294. doi: 10.1007/s11033-018-4390-6. Epub 2018 Sep 28.
Plants have many medicinal properties including anticancer activity due to the presence of several secondary metabolites. Current cancer treatment policies are not much effective because of side effects and resistance development. Therefore, the discovery of new phytotherapeutics with no or fewer side effects is highly needed. Pterospermum acerifolium (L.) wild, an angiosperm has a broad application in traditional Indian medicinal system including cancer treatment. Despite, there is no study available on the cytotoxic and apoptotic effect of P. acerifolium in human cancer cells. Exploring the medicinal properties of P. acerifolium plant by its traditional use will be helpful towards developing novel cancer therapeutics. Hence, we decided to demonstrate the anti-carcinogenic property of P. acerifolium ethanolic bark extract against lung (A549) and pancreatic (PANC-1) cancer cells. The cytotoxicity was demonstrated by MTT assay, morphological changes, and scratch invasion assay. Flow cytometry, fluorescence staining techniques, and cell cycle analysis were confirmed the apoptotic property of P. acerifolium plant. The cell viability assay revealed that P. acerifolium ethanolic bark extract significantly reduced the viability of both A549 and PANC-1 cells. Moreover, PANC-1 cells showed more sensitivity towards P. acerifolium ethanolic bark extract than A549 at higher concentrations. Clear visualization of changes such as cytoplasmic condensation, cellular morphology, cell shrinkage, and augmented number of dead cells in both the cancer cells was observed after treatment. Scratch and invasion assay showed that cell migration and invasion rate of both the cancer cells were significantly reduced. Fluorescence microscopic studies using acridine orange/ethidium bromide and DAPI (4', 6-diamidino-2-phenylindole) staining showed early and late apoptotic symptoms after treatment with bark extract. Rhodamine-123 and DCFH-DA staining analysis by fluorescence and flow cytometry showed that bark extract depolarized the mitochondria membrane potential and induced reactive oxygen species (ROS) generation. Cell cycle analysis through flow cytometry using propidium iodide stain showed that P. acerifolium bark extract arrested A549 and PANC-1 cells in sub-G1 phase stated early apoptosis. These findings collectively point to the fact that P. acerifolium bark extract induced cell cytotoxicity in lung and pancreatic cancer cells by modulating mitochondrial-mediated ROS generation, and cell cycle checkpoints.
植物具有许多药用特性，包括由于存在多种次生代谢产物而具有的抗癌活性。由于副作用和耐药性的产生，目前的癌症治疗策略效果不佳。因此，迫切需要发现副作用少或无副作用的新植物疗法。翅果油树（Pterospermum acerifolium (L.) wild），一种被子植物，在包括癌症治疗在内的印度传统医学体系中有广泛应用。尽管如此，目前尚无关于翅果油树对人类癌细胞的细胞毒性和凋亡作用的研究。通过传统用途探索翅果油树的药用特性将有助于开发新型癌症治疗方法。因此，我们决定证明翅果油树乙醇树皮提取物对肺癌（A549）和胰腺癌（PANC-1）细胞的抗癌特性。通过MTT法、形态学变化和划痕侵袭试验证明了其细胞毒性。流式细胞术、荧光染色技术和细胞周期分析证实了翅果油树的凋亡特性。细胞活力测定表明，翅果油树乙醇树皮提取物显著降低了A549和PANC-1细胞的活力。此外，在较高浓度下，PANC-1细胞对翅果油树乙醇树皮提取物的敏感性高于A549细胞。处理后，在两种癌细胞中均观察到细胞质浓缩、细胞形态、细胞收缩和死细胞数量增加等变化的清晰可视化。划痕和侵袭试验表明，两种癌细胞的细胞迁移和侵袭率均显著降低。使用吖啶橙/溴化乙锭和DAPI（4',6-二脒基-2-苯基吲哚）染色的荧光显微镜研究显示，用树皮提取物处理后出现早期和晚期凋亡症状。通过荧光和流式细胞术对罗丹明-123和DCFH-DA染色分析表明，树皮提取物使线粒体膜电位去极化并诱导活性氧（ROS）生成。使用碘化丙啶染色通过流式细胞术进行的细胞周期分析表明，翅果油树树皮提取物使A549和PANC-1细胞停滞在亚G1期，表明早期凋亡。这些发现共同表明，翅果油树树皮提取物通过调节线粒体介导的ROS生成和细胞周期检查点，诱导肺癌和胰腺癌细胞的细胞毒性。
