Hsu Shu-Chun, Lu Jih-Hung, Kuo Chao-Lin, Yang Jai-Sing, Lin Meng-Wei, Chen Guang-Wei, Su Chin-Cheng, Lu Hsu-Fung, Chung Jing-Gung
Graduate Institute of Chinese Pharmaceutical Sciences, , China Medical University, Taichung 404, Taiwan, ROC.
Anticancer Res. 2008 Mar-Apr;28(2A):1045-54.
The effects of the crude extract of Solanum lyratum (SLE) on human colon cancer colo 205 cells were investigated. The cell viability, morphological changes of the cells, cell cycle arrest, apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential (deltapsi(m)) and cell cycle- and apoptosis-associated protein levels and gene expressions were examined in colo 205 cells after exposure to various concentrations of SLE for different time periods. The results indicated that SLE decreased the percentage of viable colo 205 cells accompanied by morphological changes. The most effective concentration of SLE was 300 pg/ml (SLE 300) and this concentration was used for further investigations. SLE induced S-phase arrest and apoptosis (sub-G1) in the colo 205 cells and those effects were dose- and time-dependent. DAPI staining and DNA gel electrophoresis confirmed that SLE induced apoptosis in colo 205 cells. Flow cytometric analysis also showed that SLE 300 promoted ROS production and decreased the deltapsi(m). Western blotting analysis indicated that SLE 300 increased Bax levels and decreased Bcl-2 levels, which caused the loss of deltapsi(m) followed by cytochrome c release and caspase-9 and -3 activation, finally leading to apoptosis. SLE 300 also promoted p53 and p27, but decreased the levels of cyclin B1 thus causing S-phase arrest. The gene expression associated with those proteins was also confirmed by PCR methods. The findings show that SLE might be used as a colon cancer therapeutic agent in the future.
研究了白英粗提物（SLE）对人结肠癌colo 205细胞的作用。在不同时间段用不同浓度的SLE处理colo 205细胞后，检测细胞活力、细胞形态变化、细胞周期阻滞、凋亡、活性氧（ROS）、线粒体膜电位（ΔΨm）以及细胞周期和凋亡相关蛋白水平及基因表达。结果表明，SLE降低了存活的colo 205细胞百分比，并伴有形态变化。SLE最有效的浓度为300μg/ml（SLE 300），该浓度用于进一步研究。SLE诱导colo 205细胞发生S期阻滞和凋亡（亚G1期），且这些作用具有剂量和时间依赖性。DAPI染色和DNA凝胶电泳证实SLE诱导colo 205细胞凋亡。流式细胞术分析还显示，SLE 300促进ROS生成并降低ΔΨm。蛋白质印迹分析表明，SLE 300增加Bax水平并降低Bcl-2水平，导致ΔΨm丧失，随后细胞色素c释放以及caspase-9和-3激活，最终导致凋亡。SLE 300还促进p53和p27表达，但降低细胞周期蛋白B1水平，从而导致S期阻滞。PCR方法也证实了与这些蛋白质相关的基因表达。研究结果表明，SLE未来可能用作结肠癌治疗药物。
