Induction Study of Laminarin Sulfate LAMS-2 on Human Colon Cancer Cell Line LOVO Apoptosis through a Mitochondrial Pathway / 中国药学杂志

ABSTRACT

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. Keywords