Lee Sei-Jung, Lim Kye-Taek
Molecular Biochemistry Laboratory, Institute of Biotechnology, Chonnam National University, Kwangju, 500-757, South Korea.
Cancer Chemother Pharmacol. 2006 Apr;57(4):507-16. doi: 10.1007/s00280-005-0045-0. Epub 2005 Oct 6.
This study was carried out to investigate the apoptotic effects of glycoprotein [Solanum nigrum L. (SNL) glycoprotein, 150-kDa] isolated from Solanum nigrum L., which has been used as an antipyretic and anticancer agent in folk medicine. With the purified SNL glycoprotein, we evaluated the cytotoxic and apoptotic effects of SNL glycoprotein on HCT-116 cells, DNA fragmentation and nuclear staining assays, respectively. SNL glycoprotein has an apparent cytotoxic and apoptotic effect at a concentration of 40 microg/ml after 4 h. To further verify the apoptotic effect, we investigated the changes in activity of the apoptotic-related proteins [Bid, cytochrome c, caspases and poly(ADP-ribose)polymerase (PARP)] triggered by SNL glycoprotein, using a western blot analysis. The results in this study indicated that SNL glycoprotein has a stimulatory effect on Bid activation, resulting in the release of cytochrome c, the stimulation of caspase-8, -9 and -3 activities, and the cleavage of PARP in HCT-116 cells. However, SNL glycoprotein did not significantly stimulate an increase in levels of intracellular reactive oxygen species (ROS). From the results in this experiment, it is suggested that SNL glycoprotein induces apoptosis through the mitochondrial apoptotic signal pathway in HCT-116 cells, rather than through intracellular ROS.
本研究旨在探讨从龙葵中分离出的糖蛋白[龙葵糖蛋白,150 kDa]的凋亡作用,该糖蛋白在民间医学中被用作解热和抗癌剂。利用纯化的龙葵糖蛋白,我们分别评估了其对HCT-116细胞的细胞毒性和凋亡作用、DNA片段化和核染色试验。龙葵糖蛋白在40微克/毫升浓度下作用4小时后具有明显的细胞毒性和凋亡作用。为了进一步验证凋亡作用,我们使用蛋白质印迹分析研究了由龙葵糖蛋白触发的凋亡相关蛋白[Bid、细胞色素c、半胱天冬酶和聚(ADP-核糖)聚合酶(PARP)]活性的变化。本研究结果表明,龙葵糖蛋白对Bid激活有刺激作用,导致细胞色素c释放,刺激半胱天冬酶-8、-9和-3活性,并在HCT-116细胞中切割PARP。然而,龙葵糖蛋白并未显著刺激细胞内活性氧(ROS)水平的增加。从本实验结果来看,提示龙葵糖蛋白在HCT-116细胞中通过线粒体凋亡信号通路诱导凋亡,而非通过细胞内ROS诱导凋亡。