Apoptosis-inducing Effect of Porcine Cartilage Polysaccharide on MCF-7 Cells and Their Differentially Expressed Proteins
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Abstract:
MCF-7 (human breast adenocarcinoma cell line) cells were cultured in roller bottles to investigate the apoptosis-inducing effects of porcine cartilage polysaccharide in terms of formation of large cell aggregates. Protein expression before and after apoptosis were explored. The cell apoptotic characteristics were observed by laser scanning confocal microscopy (LSCM) after Hoechst 33342/propidium iodide (PI) staining; apoptosis was detected by Annexin V-fluorescein isothiocyanate (FITC)/PI; and changes in cell cycle were analyzed by flow cytometry (FCM). Total proteins of MCF-7 were extracted and differentially expressed proteins were analyzed by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF). The results showed that the typical apoptotic characteristics of MCF-7 cells were observed after exposure to porcine cartilage polysaccharide, and FCM results showed apoptotic rates of 20.23% and 44.7% after 24- and 48-h exposure, respectively. Additionally, it was found that the cell cycle of MCF-7 cells was arrested in S phase. Moreover, two differentially expressed proteins were identified: actin and an antioxidant protein, PRDX6.
