槐耳颗粒逆转人乳腺癌细胞MCF-7耐药的初步机制
Primary study on the mechanism of reversal drug-resistance of human breast cancer cell line MCF-7 by Huaier Granule
李戎 1谢莎 1张莉 1王先明1
作者信息
- 1. 518035,深圳市第二人民医院乳腺疾病诊疗中心
- 折叠
摘要
目的 研究槐耳颗粒逆转乳腺癌细胞株MCF-7耐药的初步机制.方法 使用四甲基偶氮唑蓝(MTT)比色法测定敏感/耐药乳腺癌细胞MCF-7-S/A对单药阿霉素(ADM)和槐耳颗粒的药物毒性,耐药倍数和槐耳颗粒对MCF-7/A的耐药逆转倍数,分别采用荧光定量逆转录-聚合酶链反应和免疫组化SP法测定多药耐药基因mdr1、多药耐药相关蛋白基因MRP的mRNA和其相应的表达产物P-gp、MRP蛋白,在MCF-7/S及非细胞毒性剂量的槐耳颗粒处理前后MCF-7/A上的表达.结果 非细胞毒性剂量(0.01 mg/ml)的槐耳颗粒能显著降低ADM对MCF-7/A的IC 50(5.06 μm),与逆转前MCF-7/A的IC50(25.8 μm),相比差异有统计学意义(P<0.01),其逆转倍数为5.1倍;0.01 mg/ml的槐耳颗粒使MCF-7/A细胞的耐药基因mdr1、MDR-1的mRNA以及相应的P-gp、MRP蛋白的表达水平均下调,与未加槐耳颗粒处理的对照组MCF-7/A相比有显著性差异(P<0.01).结论 非细胞毒性剂量槐耳颗粒具有逆转MCF-7/A细胞耐药性的作用,逆转机制和其耐药基因mdr1、MDR-1的mRNA以及相应的P-gp、MRP蛋白的表达水平下调相关,暗示槐耳颗粒是一种有潜力的耐药逆转剂.
Abstract
Objective To investigate the mechanism of the reversal drug-resistance of human breast cancer cell MCF-7 by Huaier Granule.Methods MTT method was used to detect the cell toxicity,drug-resistant multiple and reversing multiple; fluorescent quantitation reverse transcription polymerase chain reaction and immuno-histo-chemical SP staining was used to detect the mRNA level of multidrug resistance 1(mdr1),multidrug resistance-associated protein(MRP)and their corresponding proteins P-gp,MRP expression of drug sensitive cell line MCF-7/S and drug resistant cell line MCF-7/A before and after treated by non-toxicity dose of Huaier Granule,respectively.Results Non-toxicity dose(0.01 mg/ml)of Huaier Granule could significantly reduce the IC 50(5.06 μm)of ADM to the MCF-7/A,compared to the IC 50(25.8 μm)of ADM to the MCF-7/A without reverseing(P<0.01),its reversing multiple was 5.1 times.0.01 mg/ml Huaier Granule could remarkably down-regulate the mRNA level of mdr1,MRP and their corresponding proteins P-gp,MRP expression of MCF-7/A,and it showed significant reduction(P<0.01)compared to their expression of control group without Huaier Granule treatment.Conclusion Non-toxicity dose of Huaier Granule could reverse the drug-resistance of MCF-7/A,and the reversal mechanism is related to the down-regulation of the mRNA level of mdr1,MRP and their corresponding proteins P-gp,MRP expression.It implied that Huaier Granule could be a potential reversal agent for clinic.
关键词
逆转耐药/MCF-7/A/槐耳颗粒/多药耐药Key words
Drug-resistant reversal/MCF-7/A/Huaier Granule/MDR引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据