Human gastric cancer cell line SGC-7901 apoptosis induced by SFPS-B2 via a mitochondrial-mediated pathway

Biomed Mater Eng. 2014;24(1):1141-7. doi: 10.3233/BME-130914.

Abstract

This study was to investigate the effect of Sargassum fusiforme polysaccharides (SFPS-B2) on the proliferation and apoptosis of human gastric cancer cell line SGC-7901. Cells were treated with different concentrations of SFPS-B2. MTT and flow cytometry (FCM) assays were performed to evaluate the effect of SFPS-B2 on the cell growth and apoptosis. Inverted fluorescent microscope was used to observe cell morphology. Laser scanning confocal microscope (LSCM) was used to analyze intracellular calcium ion concentration, mitochondrion permeability transition pore (MPTP) and mitochondrial membrane potential (MMP). Spectrophotometer was applied to quantify the activity of Caspase-9 and Caspase-3. FCM was used to determine the expressions of Bcl-2, Bax and cytochrome C. It was shown that SFPS-B2 inhibited the growth of SGC-7901. After the treatment for 72 h, the cell apoptosis morphology was obvious, which showed that cell protuberance and apoptotic body appeared, and the cytoplasm was concentrated; the apoptotic peak appeared and the apoptotic rate increased in a dose-dependent manner. After the treatment for 24 h, SFPS-B2 activated intracellular MPTP and decreased MMP. It also increased the activity of Caspase-9 and Caspase-3, down-regulated the expression of Bcl-2 and up-regulated the expression of Bax, induced the release of Cyt-C. SFPS-B2 induced SGC-7901 apoptosis through a mitochondrial-mediated pathway, suggesting it may be an agent for cancer treatment.

Keywords: Sargassum fusiforme; apoptosis; human gastric cancer; mitochondria; polysaccharide.

MeSH terms

  • Antineoplastic Agents / pharmacology
  • Apoptosis*
  • Calcium / chemistry
  • Caspase 3 / metabolism
  • Caspase 9 / metabolism
  • Cell Line, Tumor
  • Cell Proliferation
  • Cytochromes c / metabolism
  • Cytoplasm / metabolism
  • Drug Screening Assays, Antitumor
  • Humans
  • Membrane Potential, Mitochondrial
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Mitochondria / metabolism
  • Mitochondria / pathology*
  • Polysaccharides / pharmacology*
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Sargassum / chemistry
  • Stomach Neoplasms / pathology*
  • Time Factors
  • bcl-2-Associated X Protein / metabolism

Substances

  • Antineoplastic Agents
  • BAX protein, human
  • Polysaccharides
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein
  • Cytochromes c
  • CASP3 protein, human
  • CASP9 protein, human
  • Caspase 3
  • Caspase 9
  • Calcium