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Induction Study of Laminarin Sulfate LAMS-2 on Human Colon Cancer Cell Line LOVO Apoptosis through a Mitochondrial Pathway

??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 (????m). 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 ????m; 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.