摘要
目的 观测米非司酮对卵巢癌细胞株 3AO和SKOV3细胞体外增殖和凋亡活性 ,雌激素受体 (ER)、孕激素受体 (PR)蛋白表达和细胞形态学的影响。方法 应用四甲基偶氮唑蓝 (MTT)法测定 3AO和SKOV3细胞体外增殖活性。应用流式细胞仪 (FCM )检测米非司酮在不同浓度和培养时间下 ,3AO细胞的ER、PR、p5 3和bcl 2蛋白表达率、增殖率和凋亡率。应用光学和电子显微镜观察3AO细胞的形态学变化。结果 3AO细胞在不同浓度米非司酮 (5、10、2 0、4 0、80 μmol/L)和不同时间培养 (2 4、4 8、72h)下 ,其生长抑制率由 1 7%增加至 75 0 % ,与剂量和时间呈明显正相关 (P <0 0 1) ,但SKOV3细胞增殖无明显变化 (P >0 0 5 )。 3AO细胞凋亡率与米非司酮剂量和培养时间呈正相关 (P <0 0 1)。米非司酮阻断 3AO细胞周期 ,使其停滞于G0 、G1期 ,降低S期细胞比率。米非司酮诱导 3AO细胞凋亡 ,光学和电子显微镜观察发现 ,经米非司酮培养的 3AO细胞呈现典型的细胞凋亡的特征性变化 ,包括染色体皱缩、细胞核碎裂和凋亡小体的形成。米非司酮显著升调p5 3蛋白表达 ,而降调bcl 2蛋白表达 (P <0 0 1)。米非司酮浓度为 10 μmol/L ,培养 2 4h时 ,3AO细胞 p5 3和bcl 2蛋白表达率分别为(5 4 8± 4 0 ) %和 (10 1± 1 2 ) % ,与对照的
Objective To investigate the effect of mifepristone on the activity of proliferation and the apoptosis, the expression of estrogen receptor (ER),progesterone receptor (PR) protein and morphology changes of human ovarian carcinoma cell line 3AO and SKOV3 in vitro. Methods The proliferative activity of 3AO and SKOV3, which were cultured in vitro, was measured by tetrazolium-based colorimetric assay (MTT assay). Flow cytormetry (FCM) was used to measure the expressive rate of ER, PR , p53 protein, bcl-2 protein, cell apoptotic rate and cell proliferative cycle of 3AO cells, which were cultured with different concentration and duration of mifepristone. The morphologic and ultrastructure changes of apoptotic 3AO cells was observed by the light and electron microscopy. Results Mifepristone inhibited significantly the proliferation of 3AO cells in dose-time dependent manner in vitro. The inhibitory rate of 3AO cells growth, which were cultured with different concentration of mifepristone( 5,10,20,40,80 μmol/L) and duration (24,48,72 h) was from 1.7% to 75.0%(P<0.01), but the proliferation activity of SKOV3 cells in vitro had not significant changes(P>0.05). 3AO cells apoptosis activity appeared the positive correlation with the dose of mifepristone and cultured duration (P<0.01). Mifepristone blocked 3AO cells proliferative cycle at the G 0-G 1 stage,decreased the cell radio of S stage. Mifepristone induced the apoptosis of 3AO cells in vitro. The light and electron microscopy demonstrated that 3AO cells cultured with mifepristone appeared typical morphological characteristics of apoptosis including the compaction and margination of the chromosomes, nuclear fragments and formation of apoptotic bodies. Mifepristone up-regulated significantly the expression of p53 protein, but down-regulated the expression of bcl-2 protein (P<0.01). The expressive rates of p53 and bcl-2 proteins were(54.8±4.0)% and (10.1±1.2)%, respectively, when 3AO cells was cultured with mifepristone of 10 μmol/L for 24 h. Compared with the expression rates of control group (27.1±3.3)% and (17.6±1.0)%, the difference was significant(P<0.01).The expressive rate of PR protein of 3AO cells cultured with mifepristone of 10 μmol/L for 48 h was (12.7±1.4)%,which was decreased compared with the expressive rate of control group(44.9±1.4)%(P <0.01). Conclusions Mifepristone inhibited significantly the proliferation and induced the cell apoptosis of cell line 3AO in dose-time dependent manner in vitro. The anti-tumor effect was related to down-regulation the expression of PR protein and bcl-2 protein, and to up-regulation the expression of p53 protein of 3AO cells.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2003年第10期625-628,共4页
Chinese Journal of Obstetrics and Gynecology
