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目的 考察昆布多糖硫酸酯LAMS-2对人结肠癌细胞LOVO凋亡的影响及作用途径研究.方法 采用四甲基偶氮唑蓝 (MTT) 法检测LOVO细胞的增殖,Hoechst 33258荧光染色观察细胞形态,流式细胞仪检测细胞内活性氧水平、线粒体通透性转换孔活性和线粒体膜电位,激光共聚焦显微镜观察细胞内钙离子水平,Western blot法检测凋亡蛋白Cyt-C、caspase-9和caspase-3的表达,比色法检测caspase-9和caspase-3的活性.结果 昆布多糖硫酸酯LAMS-2作用于LOVO细胞后,能显著抑制细胞的增殖,诱导细胞凋亡,细胞出现明显的凋亡特征形态,引起细胞内活性氧增多,钙离子含量增加,细胞线粒体通透性转变孔开放,线粒体膜电位降低,胞质内Cyt-C蛋白表达量增加,caspase-9和caspase-3的蛋白表达和活性都显著升高,较阴性对照组和昆布多糖组都显著增加.结论 昆布多糖硫酸酯LAMS-2可显著抑制人结肠癌LOVO细胞的增殖,并可通过线粒体途径诱导LOVO细胞发生凋亡.“,”OBJECTIVE To investigate the effects and mechanisms of LAMS-2 on the proliferation and apoptosis of human colon cancer cell line LOVO. METHODS LOVO cells were treated with different concentrations of LAMS-2, 3- ( 4, 5-Dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide ( MTT) assay and morphology observation were performed with Hoechst 33258 dye to determine the effects of LAMS-2 on the proliferation and apoptosis of LOVO cells. Flow cytometry ( FCM) was used to detect the level of reactive oxygen species ( ROS), mitochondrion permeability transition pore ( MPTP) and mitochondrial membrane potential ( △ψm). Laser scanning confocal microscope ( LSCM) was used to analyze intracellular calcium ion concentration, Western blot was performed to analyze the expressions of Cyt-C, caspase-9 and caspase-3. Spectrophotometer was applied to quantify the activity of caspase-9 and caspase-3. RESULTS LAMS-2 inhibited LOVO cell proliferation and induced apoptosis, the apoptosis morphology was obvious.LAMS-2 treatment increased the intracellular level of ROS and Ca2+; activated intracellular MPTP and decreased △ψm; it also induced the release of Cyt-C and the activation of caspase-9 and caspase-3, increased the activity of caspase-9 and caspase-3. CONCLUSION LAMS-2 can effectively inhibit the proliferation of LOVO cells and induce apoptosis through a mitochondria-mediated pathway.