五味子乙素对转染多药耐药1基因的MCF-7细胞的多药耐药逆转作用  

Effects of schisandrin B on reversing multidrug resistance in human breast cancer cells transfected with mdr1 gene

作　　者：李凌[1] 王弢[2] 许志良[3] 俞颖[3] 陈卫[4] 陈菲[5] 

机构地区：[1]浙江大学附属第二医院肿瘤研究所,杭州310009 [2]复旦大学医学院 [3]浙江省中医院 [4]浙江大学生命科学院 [5]华东师范大学生科院生物医学系

出　　处：《中华医学杂志》2005年第23期1633-1637,共5页National Medical Journal of China

摘　　要：目的探讨五味子乙素(SchB)对转染多药耐药1基因(MDR1)的人乳腺癌细胞MCF7的多药耐药逆转作用及相关机制。方法将人MDR1基因导入MCF7细胞,形成耐药细胞株MCF7/MDR1;用该细胞株为模型评价SchB的体外逆转多药耐药作用,用MTT法进行化疗药物单独或与SchB联合作用时对耐药细胞的IC50比较,计算逆转倍数。结果转染细胞MCF7/MDR表现为P糖蛋白高表达,对阿霉素、长春新碱、紫杉醇、高三尖杉酯的抗药性均增加;SchB(25μmol/L)显著减少阿霉素、长春新碱、紫杉醇和高三尖杉酯对MCF7/MDR细胞的IC50,逆转倍数达6.03～23.94倍;SchB(25μmol/L)使MCF7/MDR细胞对若丹明123的胞内积聚增加约5倍,效果与维拉帕米10μmol/L浓度时相当;但SchB(25μmol/L)不影响MCF7/MDR细胞的P糖蛋白表达。结论SchB能有效逆转转染MDR1的MCF7细胞的多药耐药,其机制可能是抑制了P糖蛋白的药物外排生物学活性。Objective To investigate the multidrug resistance (MDR) reversal activity of schisandrin B (SchB) in transfected human breast cancer cell line MCF-7/MDR1.Methods Human breast cancer cells of the line MCF-7 were cultured and transfected with mdr1 gene so as to establish a P-glycoprotein (P-gp) stable-expressing cell line MCF-7/MDR1. The expression of P-gp in the MCF-7/mdr1 cells was assayed by flow cytometry using fluorescent antibody. MCF-7 cells transfected with blank plasmid MCF-7/neo was used as controls. Adriamycin (ADR), verapamil (VER), pacilitaxel (taxol), and homoharringtonine (HHT) were added into the culture fluid of the MCF-7/mdr1 cells and MTT method was used to detect the IC 50 of these drugs. The culture fluid of the MCF-7/MDR1 cells was added with SchB of the concentrations of 2.5, 12.5, 25, and 50 μmol/L and then added with ADR1 cells, MTT method was used to calculate the reversal effect (RF) of SchB on the MDR phenotype of the MCF-7/mdr1 cells. MTT method was used to calculate the RF. Another culture fluid of MCF-7/mdr1 cells was added with 25 μmol/L SchB and then with ADR, vincristine (VCR), pacilitaxel, and HHT with that added with 10μmol/L VER as control. After treatment with SchB the MF7/mgr1 cells were co-incubated with 5 μmol/L rhodamine (Rh)-123, then flow cytometry was used to detect the accumulation of Rh-123 within the cells. After treatment with 25 μmol/L SchB for 0, 0.5, 1, 6, 24, 48, and 72 hours flow cytometry was used to detect the P-gp expression in the MF7/mgr1 cells. Results The transfected MCF-7/MDR1 cells overexpressed P-gp and exhibited resistance to multiple drugs, including ADR,VCR, pacilitaxel and HHT. SchB (25 μmol/L) significantly enhanced the sensitivity of the MCF-7/MDR1 cells to above mentioned chemotherapeutic agents, with a reversal factors of 6.03 to 23.94 times. The effect of SchB (25 μmol/L) on Rh-123 accumulation in MDR cells was equivalent to that of 10 μmol/L VER, however no significant difference was found in the effect of SchB (25 μmol/L) on the

关 键 词：五味子乙素 转染多药耐药1基因 MCF-7细胞 多药耐药 逆转作用 中药 

分 类 号：R285[医药卫生—中药学]