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目的:探寻大环内酯类药物多拉菌素逆转人乳腺癌多药耐药细胞株(MCF-7/adr)多药耐药(multidrug resistance,MDR)的作用及机制。方法:采用MTT比色法测定细胞生长抑制率及逆转倍数;高效液相色谱(HPLC)法检测细胞内阿霉素(ADR)的积累变化;荧光分光光度仪检测罗丹明123(Rh123)在肿瘤细胞内的积累与外排;通过RT-PCR与流式细胞仪检测MDR1基因与P-糖蛋白(P-gp)表达的变化。结果:5μmol·L-1的多拉菌素可明显增强MCF-7/adr对ADR的敏感性,逆转倍数可达28.93,并增强了细胞内ADR及Rh123的积累,呈剂量依赖关系。同时,MDR1及P-gp的表达降低。结论:多拉菌素对MCF-7/adr的耐药有一定的逆转作用,有希望成为肿瘤多药耐药的逆转剂。
OBJECTIVE: To investigate the effect and mechanism of macrolide drug on reversing multidrug resistance (MDR) in human breast cancer multidrug resistance cell line (MCF-7 / adr). Methods: MTT colorimetric method was used to determine the cell growth inhibitory rate and reversal fold. The accumulation of doxorubicin (ADR) was detected by high performance liquid chromatography (HPLC). Fluorescence spectrophotometry was used to detect Rh123 in the tumor Cell accumulation and efflux. The changes of MDR1 gene and P-glycoprotein (P-gp) expression were detected by RT-PCR and flow cytometry. RESULTS: Doxorubicin at 5μmol·L-1 significantly increased the sensitivity of MCF-7 / adr to ADR. The reversal factor was up to 28.93 and the accumulation of ADR and Rh123 was increased in a dose-dependent manner. At the same time, the expression of MDR1 and P-gp decreased. Conclusion: Doramectin can reverse the drug resistance of MCF-7 / adr and hopefully become a reversal agent for MDR.