University of Baghdad, College of Science, Department of Biology, Baghdad, Iraq.
University of Karachi, International Center for Chemical and Biological Sciences, Dr. Panjwani center for Molecular Medicine and Drug Research, Karachi, Pakistan.
J Appl Biomed. 2021 Mar;19(1):40-47. doi: 10.32725/jab.2021.007. Epub 2021 Mar 1.
Clausena excavata Burm. f is a plant used in folklore medicine for the treatment of various ailments in South East Asia. The plant parts contain chemical components that are cytotoxic to many cancer cells.
The study investigated the cytotoxic effects of ethyl acetate, methanol and chloroform C. excavata leaf extracts on the non-small-lung cancer, NCI-H460, cell line.
Based on the 3-(4,5-dimethylthiazol-2-yl)-2,5,-diphenyltetrazolium bromide (MTT) assay, among extracts, ethyl acetate C. excavata leaf extract (EACE) was the most potent anti-NCI-H460 cells, with IC50 value of 47.1 ± 6.1 μg/ml. The effects of EACE on NCI-H460 cells were also determined by clonogenic, 4', 6-diamidino-2-phenylindole (DAPI), and annexin-V-fluorescein isothiocyanate/propidium iodide-PI flow cytometric assays. Reactive oxygen species (ROS) production and apoptotic gene expressions was determined via flow cytometry and real-time quantitative PCR, respectively.
EACE-treated NCI-H460 cells after 48 h underwent apoptosis as evident by loss of cell viability, cell shrinkage, and chromatin condensation. The results also showed EACE mediated increase in ROS production by the NCI-H460 cells. After 48 h treatment, EACE increased the pro-apoptotic BAX and decreased the anti-apoptotic Bcl-2, Survivin and c-Myc gene expressions.
EACE is a potential anti-lung cancer by increasing cancer cell ROS production and apoptosis.
山胡椒（ Clausena excavata Burm. f）是一种在东南亚民间医学中用于治疗各种疾病的植物。植物部分含有对许多癌细胞具有细胞毒性的化学成分。
本研究调查了乙酸乙酯、甲醇和氯仿 C. excavata 叶提取物对非小细胞肺癌 NCI-H460 细胞系的细胞毒性作用。
基于 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法，在提取物中，乙酸乙酯 C. excavata 叶提取物（EACE）对 NCI-H460 细胞的抑制作用最强，IC50 值为 47.1±6.1μg/ml。EACE 对 NCI-H460 细胞的作用也通过集落形成、4'，6-二脒基-2-苯基吲哚（DAPI）和 Annexin-V-荧光素异硫氰酸酯/碘化丙啶-PI 流式细胞术测定法来确定。通过流式细胞术和实时定量 PCR 分别测定活性氧（ROS）的产生和凋亡基因的表达。
EACE 处理 48 h 后的 NCI-H460 细胞发生凋亡，表现为细胞活力丧失、细胞皱缩和染色质浓缩。结果还表明，EACE 介导的 NCI-H460 细胞中 ROS 产生增加。经过 48 h 处理，EACE 增加了促凋亡 BAX 的表达，降低了抗凋亡 Bcl-2、Survivin 和 c-Myc 基因的表达。
EACE 通过增加癌细胞 ROS 产生和凋亡，是一种有潜力的肺癌治疗方法。
