Research on the mechanisms of streptococcal protein inducing cycle arrest and apoptosis in glioblastoma BT325 cell

doi:10.3969/j.issn.1673-8640.2014.04.015

Abstract

Abstract:

Objective　To investigate the mechanisms of streptococcal protein inducing cycle arrest and apoptosis in human cerebral glioblastoma BT325 cell. Methods　Human cerebral glioblastoma BT325 cells were cultured in vitro， and methlcyclopentadienyl manganese tricarbonyl（MTT） assay was used to determine the proliferation activity of BT325 cells. The morphological changes of BT325 cells were observed by the phase-contrast microscopy. The apoptotic cells were observed after fluorescence staining. Flow cytometry was used to determine the cell cycle， the rate of cell apoptosis and the change of mitochondrial membrane potential（MMP）. The expression of apoptosis-related proteins was determined by Western blot. Results　Streptococcal protein（10-100 mg/L） inhibited the proliferation of BT325 cells significantly and induced G2/M phase arrest in time- and dose-dependent manners. MMP declined（P＜0.01）， and apoptotic cells gradually increased after BT325 cells were treated with streptococcal protein for 24 h. The rate of cell apoptosis in determination group was significantly higher than that in control group at 48h（P＜0.01）. In a time-dependent manner， streptococcal protein（50 mg/L） up-regulated the expression of cell P53 protein， down-regulated the expression of B-cell lymphoma-2（Bcl-2）， led to the release of mitochondrial cytochrome C（CytC） from mitochondria into cytoplasm， and then caused the activation of the precursor of cysteinyl aspartate specific proteinase 3（Procaspase-3）. Conclusions　Streptococcal protein may exert its anticancer activity through inhibiting the proliferation of human cerebral glioblastoma BT325 cells， effecting the distribution of cell cycle and inducing cell apoptosis by mitochondrial-dependent pathway.

Key words: Streptococcus, Human cerebral glioblastoma BT325 cell, Cell cycle, Cell apoptosis, Mitochondrial membrane potential

CLC Number:

R446.62

WANG Yan，ZHANG Liping，CHEN Hui，CAO Ning，ZHU Junping，PING Guoling，LI Weihong. Research on the mechanisms of streptococcal protein inducing cycle arrest and apoptosis in glioblastoma BT325 cell[J]. , 2014, 29(4): 363-368.

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