米非司酮对耐药卵巢癌细胞增殖、凋亡及其对顺铂敏感性的影响

Effects of Mifepristone on the Proliferation,Apoptosis,and Cis-Platinum (DDP) Sensitivity of Chemo-resistant Human Ovarian Cancer Cells

背景与目的:米非司酮是有效的孕酮受体拮抗剂。研究发现,米非司酮对体内外卵巢癌细胞均具有生长抑制作用,但其机制尚不清楚。本研究的目的在于探讨米非司酮对耐药卵巢癌细胞增殖、凋亡及对顺铂(DDP)敏感性的影响和机制,为临床应用米非司酮治疗难治性卵巢癌提供实验依据。方法:体外培养耐药卵巢癌细王刚,等.米非司酮对耐药卵巢癌细胞株SK-OV-3,采用MTT法检测单用米非司酮及合用顺铂时对SK-OV-3细胞增殖的影响,分析米非司酮与顺铂在抑制耐药卵巢癌细胞增殖中的相互作用。采用终末脱氧核糖核苷酸转移酶介导的原位缺口末端标记法和流式细胞术,分析米非司酮及米非司酮联合顺铂对卵巢癌细胞周期和凋亡的影响。结果:实验所选各种浓度米非司酮对SK-OV-3细胞均表现出一定程度的生长抑制作用,并呈现浓度依赖性。当顺铂浓度为1.25μg/ml或2.5μg/ml,合用米非司酮浓度为20μg/ml、10μg/ml、5μg/ml、2.5μg/ml、1.25μg/ml或0.625μg/ml时,能显著抑制SK-OV-3细胞的增殖,并显示出两种药物的协同作用(q>1.15)。当顺铂浓度为0.625μg/ml或5.0μg/ml时,仅表现为两种药物的相加作用(0.85<q<1.15)。米非司酮可诱导SK-OV-3细胞凋亡,并将细胞阻滞于G0/G1期。当米非司酮浓度为1.25μg/ml、2.50μg/ml和5.00μg/ml时。

BACKGROUND & OBJECTIVE: Mifepristone is an effective progeste ro ne receptor antagonist. It was reported that mifepristone can inhibit the growth of ovarian carcinoma cells either in vitro or in vivo, but the exact mechanism is unknown. The effect of mifepristone on the growth, apoptosis and cis- platin um (DDP)- sensitivity of chemo- resistant ovarian carcinoma cell lines have sc arcely been reported .The purpose of this study was to investigate the effect an d its mechanism of mifepristone on the proliferation, apoptosis and DDP sensitiv ity of DDP- resistant human ovarian carcinoma cells, and to give experimental b asis for treating refractory ovarian carcinoma with mifepristone. METHODS: DDP- resistant human ovarian carcinoma cell line SK- OV- 3 cells were cultured in vitro, and the MTT assay was used to examine the antiproliferative effect of mif epristone with or without DDP on SK- OV- 3 cells. The cooperative effects betw een mifepristone and DDP in inhibiting the growth of SK- OV- 3 cells were anal yzed. TdT mediated dUTP nick end labeling (TUNEL) and flow cytometry (FCM) were used to examine the effects of mifepristone with or without DDP on the apoptosis and cell cycle of SK- OV- 3 cells.RESULTS: Mifepristone produced concentratio n- dependent antiproliferative effect on SK- OV- 3 cells at all experimental concentrations.Enhanced antiproliferative effects were found when SK- OV- 3 ce lls were cultured with mifepristone at 0.625, 1.25, 2.5, 5, 10, and 20 μ g/ml c ombined with 1.25 μ g/ml or 2.5 μ g/ml DDP (q >1.15). Only additive effects we re found when the cells were cultured with mifepristone and 0.625 μ g/ml or 5.0 μ g/ml DDP (0.85< q< 1.15). Mifepristone induced concentration- dependent apo ptosis in SK- OV- 3 cells and arrested cells in the G0/G1- phase of cell cycl e. The apoptosis rate were 14.52% , 36.14% , and 53.22% ,respectively,when th e cells were cultured with mifepristone at 1.25, 2.50, and 5.00 μ g/ml. The per centage of G0/G1- phase cells was increased with the concentration of mifeprist one. Synergic effect between mifepristone (at 1.25, 2.5, and 5 μ g/ml) and 2.5 μ g/ml DDP was found in inducing SK- OV- 3 cells apoptosis (q >1.15) and G0/G 1- phase stasis. CONCLUSION: Mifepritstone can inhibit the growth of chemo- re sistant human ovarian carcinoma cells,and enhance its DDP sensitivity. This may be associated with the synergic effect between mifepristone and DDP in inducing apoptosis and G0/G1- phase stasis.