Engineering Research Center of Natural Anticancer Drugs, Ministry of Education, Harbin University of Commerce, Harbin 150076, China.
Molecules. 2012 Aug 20;17(8):9947-60. doi: 10.3390/molecules17089947.
Many scientific studies have shown that laminarin has anti-tumor effects, but the anti-tumor mechanism was unclear. The purpose of this study was to investigate the effect of laminarin on the induction of apoptosis in human colon cancer LOVO cells and the molecular mechanism involved. LOVO cells were treated with different concentrations of laminarin at different times. Morphology observations were performed to determine the effects of laminarin on apoptosis of LOVO cells. Flow cytometry (FCM) was used to detect the level of intracellular reactive oxygen species (ROS) and pH. Laser scanning confocal microscope (LSCM) was used to analyze intracellular calcium ion concentration, mitochondrion permeability transition pore (MPTP) and mitochondrial membrane potential (MMP). Western blotd were performed to analyze the expressions of Cyt-C, Caspase-9 and -3. The results showed the apoptosis morphology, which showed cell protuberance, concentrated cytoplasm and apoptotic bodies, was obvious after 72 h treatment. Laminarin treatment for 24 h increased the intracellular level of ROS and Ca²⁺; decreased pH value; activated intracellular MPTP and decreased MMP in dose-dependent manners. It also induced the release of Cyt-C and the activation of Caspase-9 and -3. In conclusion, laminarin induces LOVO cell apoptosis through a mitochondrial pathway, suggesting that it could be a potent agent for cancer prevention and treatment.
许多科学研究表明,昆布多糖具有抗肿瘤作用,但抗肿瘤机制尚不清楚。本研究旨在探讨昆布多糖对人结肠癌细胞 LOVO 细胞凋亡的诱导作用及其涉及的分子机制。用不同浓度的昆布多糖分别处理 LOVO 细胞不同时间。观察细胞形态学变化,确定昆布多糖对 LOVO 细胞凋亡的影响。采用流式细胞术(FCM)检测细胞内活性氧(ROS)和 pH 值水平。激光扫描共聚焦显微镜(LSCM)分析细胞内钙离子浓度、线粒体通透性转换孔(MPTP)和线粒体膜电位(MMP)。Western blot 分析 Cyt-C、Caspase-9 和 -3 的表达。结果显示,72 h 处理后,细胞突起、细胞质浓缩和凋亡小体等凋亡形态明显。昆布多糖处理 24 h 后,细胞内 ROS 和 Ca²⁺水平升高,pH 值降低,MPTP 激活,MMP 降低,呈剂量依赖性。还诱导 Cyt-C 释放和 Caspase-9 和 -3 的激活。综上所述,昆布多糖通过线粒体途径诱导 LOVO 细胞凋亡,提示其可能是一种有效的癌症预防和治疗药物。