解毒祛瘀方对乳腺癌细胞MCF-7/ADM的耐药逆转作用

Research on Jiedu Quyu Prescription in Drug Resistance Reversal in MCF-7/ADM Human Breast Cancer Cells

目的：研究解毒祛瘀方对人乳腺癌耐药细胞MCF-7/ADM耐药逆转的作用及机制。方法：以MCF-7/ADM细胞为研究对象,利用噻唑蓝（MTT）比色法检测解毒祛瘀方对人乳腺癌耐药细胞MCF-7/ADM生长的影响;应用流式细胞术检测肿瘤细胞内罗丹明123（Rh-123）的含量;分别利用实时荧光定量聚合酶链式反应（Real-time PCR）及蛋白免疫印迹法（Western blot）检测肿瘤细胞内多药耐药蛋白1（MDR1）,乳腺癌耐药相关蛋白（BCRP）mRNA及蛋白表达变化。结果：与空白组比较,经1.25,2.5 g·L^-1解毒祛瘀方作用后,阿霉素对人乳腺癌耐药细胞MCF-7/ADM的逆转倍数（RF）分别提升1.7倍和3.0倍（P〈0.05）;人乳腺癌耐药细胞MCF-7/ADM中Rh-123含量分别提高了1.8倍和2.5倍（P〈0.05）,MDR1和BCRP蛋白和mRNA表达水平明显下降（P〈0.05）,1.25,2.5 g·L^-1解毒祛瘀方MDR1 mRNA表达分别降低35.5%和56.0%（P〈0.05）,BCRP mRNA表达分别降低41.6%和49.5%（P〈0.05）。结论：解毒祛瘀方可提高人乳腺癌耐药细胞MCF-7/ADM对阿霉素的敏感性,逆转该细胞对阿霉素的耐药性,其机制可能与降低MDR1和BCRP蛋白和mRNA的表达,抑制细胞药物外排作用相关。

Objective： To investigate the effect and mechanism of Jiedu Quyu prescription on drug resistance reversal for MCF-7/ADM human breast cancer cells. Method： The 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） was performed to test the effect of Jiedu Quyu prescription on proliferation of MCF-7/ADM cells. Flow cytometry was used to detect the concentration of intracellular rhodamine（Rh）-123,Real-time PCR and Western blot were used to analyze the expression of genes and proteins,including multiple drug resistant 1（MDR1） and breast cancer resistant protein（BCRP）. Result： After treatment with 1. 25,2. 50 g·L^-1 Jiedu Quyu prescription,the reverse fold（RF） of MCF-7/ADM was 1. 7 and 3. 0 respectively（P〈0. 05）; the concentration of intracellular Rh-123 was enhance for 1. 8 and 2. 5 times respectively as compared with control group（P〈0. 05）. The protein and mRNA expression levels of MDR1 and BCRP were down-regulated obviously（P〈0. 05）; MDR1 mRNA expression level was decreased by 35. 5% and 56. 0% respectively（P〈0. 05）,while BCRP mRNA expression level was decreased by 41. 6% and 49. 5% respectively（P〈0. 05）. Conclusion：Jiedu Quyu prescription could enhance the sensitivity of human breast cancer cell line MCF-7/ADM to adriamycin,and reverse drug resistance of MCF-7/ADM. Its mechanism may be correlated with down-regulation of MDR1 and BCRP,as well as inhibition of intracellular drug efflux.