The Study Of MCF-7 Cultured In Roller Bottle And Cartilage Polysaccharide-Induced Apoptosis In MCF-7

The large-scale culture of MCF-7, a kind of human breast cancer cells, was studied by roller bottle on the basis of MCF-7 flask cultured. Then the apoptotic effect and mechanism of cartilage polysaccharide on MCF-7 in roller bottle was also exlpored.Firstly the ranges of roller bottle early stage rotation rate and adherent rotation rate were determined respectively by univariate experiment. Then the inoculation amount, medium amount and culture time were evaluated by orthogonal experiment of three levels and three fators. The rusults indicated the production of cells was affected significantly mainly by the fators of inoculation amount and culture time. The optimization conditions for MCF-7 roller bottle culture were:early rotation rate 15r/h, adherent rotation rate 40r/h, inoculation amount 5×10 cells, culture time 96h, medium amount 150mL.The apoptosis effect of cartilage polysaccharide on the MCF-7 cells was researched after roller bottle cultivation was established. The typical apoptosis characteristics of MCF-7 cells induced by cartilage polysaccharide were observed by Hochest33342/PI double staining, Annexin V-FITC/PI and DNA Ladder. The apoptosis rate was 20.23% and 44.7% respectively by flow cytometry after MCF-7 was cultured with cartilage polysaccharide for 24h and 48h. The cell cycle was arrested in S phase.Total protein maps were analysed between Normal MCF-7 and MCF-7 cultured with cartilage polysaccharide by two-dimensional electrophoresis technology.10 proteins expressed differentially were detected,7 of those were down-expressed and 3 of those were up-expressed.3 points selected as the differentially expressed, but only 2 proteins were identified by MALDI-TOF-MS/MS. One was actin which was down-expressed to induce mitochondrial apoptosis. the orher was PRDX6, a member of antioxidant protein family, which was down-expressed to result in oxidative injury of cells apoptosis.