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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)的表达变化。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蛋白表达水平明显下降(P<0.05),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蛋白的表达,抑制细胞药物外排作用相关。
Objective: To study the effect and mechanism of Jiedu Quyu on the reversal of multidrug resistance in human breast cancer cell line MCF-7 / ADM. Methods: MCF-7 / ADM cells were used as experimental materials, the effect of Jiedu Quyu prescription on the growth of human breast cancer cell line MCF-7 / ADM was detected by MTT colorimetric assay. Flow cytometry was used to detect tumor Intracellular Rhodamine 123 (Rh-123) was detected by real-time PCR and Western blot respectively. The expression of MDR1 ), Breast cancer resistance-related protein (BCRP) expression changes. Results: Compared with the blank group, the reversal times (RFs) of doxorubicin on human breast cancer cell line MCF-7 / ADM were increased by 1.7, 2.5 g · L ~ (-1) (P <0.05). The content of Rh-123 in MCF-7 / ADM cells increased by 1.8 times and 2.5 times (P <0.05), while the expression of MDR1 and BCRP protein decreased <0.05). The mRNA expression of MDR1 decreased by 35.5% and 56.0% (P <0.05), while the expression of BCRP mRNA decreased by 41.6% and 49.5% (P <0.05), respectively. Conclusion: Jiedu Quyu can increase the sensitivity of human breast cancer cell line MCF-7 / ADM to doxorubicin and reverse the drug resistance to doxorubicin, which may be related to the decrease of MDR1 and BCRP protein expression , Inhibition of cell drug efflux related.