Li Qiang, Li Ji-jun, Zhao Xing-bo, Ji Mei
Department of Obstetrics and Gynecology, Shandong Provincial Hospital, Jinan 250021, China.
Zhonghua Fu Chan Ke Za Zhi. 2003 Oct;38(10):625-8.
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.
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.
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 micro mol/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 micro mol/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 micro mol/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).
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.
探讨米非司酮对人卵巢癌细胞系3AO和SKOV3体外增殖活性、凋亡、雌激素受体（ER）、孕激素受体（PR）蛋白表达及形态学变化的影响。
采用四氮唑盐比色法（MTT法）检测体外培养的3AO和SKOV3的增殖活性。用流式细胞术（FCM）检测不同浓度和作用时间米非司酮作用后3AO细胞的ER、PR、p53蛋白、bcl-2蛋白表达率、细胞凋亡率及细胞增殖周期。用光镜和电镜观察凋亡3AO细胞的形态及超微结构变化。
米非司酮体外能明显抑制3AO细胞增殖，呈剂量-时间依赖性。不同浓度（5、10、20、40、80 μmol/L）和作用时间（24、48、72 h）米非司酮作用后3AO细胞生长抑制率为1.7%～75.0%（P<0.01），而对SKOV3细胞体外增殖活性无明显影响（P>0.05）。3AO细胞凋亡活性与米非司酮剂量和作用时间呈正相关（P<0.01）。米非司酮将3AO细胞增殖周期阻断于G(0)-G(1)期，使S期细胞比例下降。米非司酮体外诱导3AO细胞凋亡。光镜和电镜显示米非司酮作用后的3AO细胞呈现典型的凋亡形态学特征，包括染色体固缩、边缘化、核碎片及凋亡小体形成。米非司酮明显上调p53蛋白表达，下调bcl-2蛋白表达（P<0.01）。3AO细胞用10 μmol/L米非司酮作用24 h后，p53和bcl-2蛋白表达率分别为(54.8±4.0)%和(10.1±1.2)%，与对照组表达率(27.1±3.3)%和(17.6±1.0)%相比，差异有统计学意义（P<0.01）。3AO细胞用10 μmol/L米非司酮作用48 h后PR蛋白表达率为(12.7±1.4)%，较对照组表达率(44.9±1.4)%降低（P<0.01）。
米非司酮体外能明显抑制3AO细胞增殖并诱导其凋亡，呈剂量-时间依赖性。其抗肿瘤作用与下调3AO细胞PR蛋白和bcl-2蛋白表达及上调p53蛋白表达有关。
