Zhu Fu-Rong, Li Yong-Ning, He Shu-Lan, Chen Qian-Shun, Xu Xun-Yu
Department of Pharmacy, Fujian Health College, Fuzhou, 350101, China.
Department of Thoracic Surgery, Fujian Provincial Hospital, Provincial Clinical College of Fujian Medical University, Fuzhou, 350001, China.
Chin J Integr Med. 2018 Oct;24(10):763-767. doi: 10.1007/s11655-018-2839-z. Epub 2018 Jun 21.
To investigate the anti-proliferative effects of saponins prepared from Plena Clematis (PC) cultured in Fujian Province, China on 4 human tumor cell lines and its possible anti-tumor mechanism.
The growth inhibition assays of saponins on human esophageal squamous carcinoma cell line (EC9706), human hepatoma cell line (HepG-2), human oral cancer cell line (KB) and human gastric cancer cell line (BGC-823) were evaluated in vitro by thiazolyl blue (MTT) method. The inhibitory effects on EC9706 treated with different concentrations of saponins (15.62, 31.25, 62.50, 125, 250 and 500 μg/mL) were performed in vitro by MTT method. The morphology and nuclear staining with acridine orange/ethidium bromide of EC9706 cells treated with saponins were illustrated under an inverted phase fluorescence microscope. The apoptotic effects of saponins were further evaluated by annexin-V/propidium iodide dual staining experiment to examine the occurrence of phosphatidylserine externalization onto the cell surface by a flflow cytometer.
MTT assay showed that the saponins could inhibit the proliferation of 4 tumor cell lines. Among them, the maximum inhibition rate of 73.1% was detected in EC9706 cells at the saponins concentration of 250 μg/mL for 24 h. Further investigation indicated that the saponins induced EC9706 cells apoposis. The EC9706 cells presented apoptotic characteristics when treated with saponins, including that the morphologies of EC9706 cells were appeared round-shaped with higher refraction, and the cell nuclear stained orange with EB after 250 μg/mL saponins exposure. The flow cytometry analysis results showed that the induction of cell cycle arrest in apoptotic system may participate in the anti-proliferative activity of saponins on EC9706 cells.
The saponins from PC exhibited significant cytotoxicity against human EC9706, KB, BGC-823, and HepG-2 cells and might be beneficial to development of ethnic pharmaceutical plant for potential anti-tumor drugs.
研究中国福建省栽培的威灵仙皂苷对4种人肿瘤细胞系的抗增殖作用及其可能的抗肿瘤机制。
采用噻唑蓝（MTT）法体外评价皂苷对人食管鳞癌细胞系（EC9706）、人肝癌细胞系（HepG-2）、人口腔癌细胞系（KB）和人胃癌细胞系（BGC-823）的生长抑制作用。用不同浓度（15.62、31.25、62.50、125、250和500μg/mL）的皂苷处理EC9706细胞，采用MTT法体外检测其抑制作用。在倒置相差荧光显微镜下观察皂苷处理的EC9706细胞的形态及吖啶橙/溴化乙锭核染色情况。通过膜联蛋白V/碘化丙啶双染实验进一步评价皂苷的凋亡作用，用流式细胞仪检测细胞表面磷脂酰丝氨酸外翻的发生情况。
MTT法检测结果显示，皂苷能抑制4种肿瘤细胞系的增殖。其中，在皂苷浓度为250μg/mL处理24 h时，EC9706细胞的最大抑制率为73.1%。进一步研究表明，皂苷可诱导EC9706细胞凋亡。皂苷处理后的EC9706细胞呈现凋亡特征，包括细胞形态呈圆形，折光性增强，250μg/mL皂苷处理后细胞核被EB染成橙色。流式细胞术分析结果表明，凋亡系统中细胞周期阻滞的诱导可能参与了皂苷对EC9706细胞的抗增殖活性。
威灵仙皂苷对人EC9706、KB、BGC-823和HepG-2细胞具有显著的细胞毒性，可能有利于民族药用植物开发潜在的抗肿瘤药物。
