诺美孕酮逆转乳腺癌耐药细胞株多药耐药性的研究  

Reversal of nomegestrol acetate on multidrug resistance in drug resistant human breast cancer cell line MCF7/ADR

作　　者：李杰[1] 许良中[2] 何开玲[2] 郭伟剑[1] 朱雄增[2] 郑云红[3] 夏鹏[2] 

机构地区：[1]上海第二医科大学附属新华医院肿瘤中心肿瘤科,200092 [2]上海医科大学 [3]中国科学院上海药物研究所

出　　处：《中华肿瘤杂志》2002年第2期129-132,共4页Chinese Journal of Oncology

摘　　要：目的　研究诺美孕酮 (NOM)对乳腺癌耐药细胞株 (MCF7/ADR)多药耐药性 (MDR)的影响 ,探讨其调节机制。方法　应用MTT法 ,研究NOM对MCF7/ADR药物敏感性的影响。采用半定量逆转录聚合酶联反应 (RT PCR)和免疫细胞化学染色 ,分析耐药基因MDR1、谷胱甘肽S转移酶Pi(GSTπ)、拓扑异构酶Ⅱα(TopoⅡα)和多药耐药相关蛋白 (MRP)表达的变化。通过流式细胞技术观察NOM对MCF7/ADR细胞内药物积累和细胞周期的影响。结果　NOM对MCF7/ADR的MDR具有明显的逆转作用 ,在 2 0 ,10和 5 μmol/L浓度时的逆转倍数分别为 2 1,12和 8倍 ,逆转强度明显高于前身化合物甲地孕酮 ,而与维拉帕米相当。 5 μmol/LNOM处理MCF7/ADR后 ,MDR1的mRNA表达水平明显降低 ,呈现时间依赖性变化 ;P糖蛋白 (P gp)和GSTπ蛋白的变化符合相应mRNA表达的变化 ;MRP和TopoⅡα基因表达未见明显变化。用NOM 2 0 ,10和 5 μmol/L分别处理 2h后 ,ADR在细胞内的积累分别增加到 2 .7倍、2 .3倍和 1.5倍。同时 ,NOM可明显加强ADR对MCF7/ADR细胞在G2 M期的阻滞作用。结论　NOM具有较强的逆转MCF7/ADR细胞MDR的作用 ,其逆转机制为多种途径 ,包括时间依赖性下调MDRI和GSTπ基因的表达 ,增加细胞内药物积累 ,加强ADR对MCF7/ADR在G2Objective To study the reversal effect of nomegestrol acetate (NOM) on mutidrug resistanc e (MDR) in MCF7/ADR and its mechanism. Methods Using tetrazolium dye assay, effects of various concentrations of NOM on sen sitivity to ADR in MCF7/ADR was studied. Expression of MDR related genes MDR1, g lutathoine S transferase Pi (GSTπ), Topoisomerase Ⅱα (TopoⅡα) and MDR rela ted protein (MRP) were assayed by reverse transcription polymerase chain reacti on (RT PCR) and immunocytochemistry assay. Using flow cytometry (FCM), intrace llular ADR concentration effects on cell cycle were observed. Results NOM significantly reversed MDR in MCF7/ADR. After NOM 20, 10 and 5 μmol/L treatment, the chemosensitivity to ADR increased to 21, 12 and 8 times. Th e reversal activity of NOM was stronger than that of the precursor compound meg estrol acetate, and was comparable to that of verapamail. After treatment w ith NOM 5 μmol/L both MDR1 and GSTπ mRNA genes expression began to decline on D2 ( P <0.05, & P <0.01) and reached the lowest leve l on D3 (both P <0.01), but the expression levels began to rise on D6 again (both P <0.05). The expression of MRP and TopoⅡα gave n o significant change. Changes of P gp and GSTπ protein expressions were simila r to those of their mRNA expressions, showing early decline and late rise. T wo hours after NOM 20, 10, and 5 μmol/L treatment, intracellular ADR concentrat ion increased 2.7, 2.3 and 1.5 times, respectively. FCM data showed that after forty eight hours, combined administration of NOM ( 2 0 μmol/L) and ADR (from low concentration to high concentration), MCF7/ADR cell s showed gradual arrest in the G 2M phase with the increase of ADR dose. Conclusion NOM has strong reversal effects on MDR in MCF7/AD R. The reversal takes place via different routes, i.e. down regulating mRNA and protein expression levels of MDR1 and GSTπ, increasing intracellular drug concentration , and enhancing the arrest of ADR in cells at G 2M phase.

关 键 词：诺美孕酮 多药耐药 乳腺癌 耐药细胞株 

分 类 号：R737.9[医药卫生—肿瘤]