葡萄籽多酚逆转人乳腺癌多药耐药性及其机制的研究

Grape seed polyphenols reverses multidrug resistance of human breast cancer

目的探讨葡萄籽多酚 (grapeseedpolyphenols ,GSP)逆转肿瘤多药耐药的作用及其相关机制。方法以人乳腺癌MCF 7细胞及其耐药株MCF 7/ADR细胞作为实验对象 ,用MTT法观察GSP对MDR的逆转作用 ,采用Westernblot、Northernblot检测GSP对MDR1基因表达的影响 ;流式细胞术 (FCM)观察GSP对P 糖蛋白 (P gp)功能的影响。结果非细胞毒性剂量GSP 1 2mg/L、2 4mg/L均能逆转MCF 7/ADR细胞的多药耐药性 ,逆转倍数分别为 5 77和 9 79倍 ;1 2mg/L、2 4mg/LGSP分别使P gp表达降至对照组的 80 83% (t=5 5 8,P <0 0 5 )和 5 0 81% (t =10 96 ,P <0 0 1) ;1 2mg/L、2 4mg/LGSP下调MDR1mRNA表达 ,降至对照组的 6 0 92 % (t =16 0 3,P <0 0 0 5 )和 4 2 0 5 % (t =2 1 70 ,P <0 0 0 5 ) ;2 4mg/LGSP可增加细胞内阿霉素 (ADM)的积累。结论体外试验证实GSP具有多药耐药逆转作用 ,是一种潜在的化疗增敏剂。

Objective To explore if grape seed polyphenols (GSP) reverses human breast cancer multidrug resistance. MethodsIn this study,adriamycin-resistant human breast carcinoma cells ( MCF-7/ADR ) and its parental cells (MCF-7) were used to determine the effect of GSP. MTT assay was adopted to evaluate the cytotoxity. Western blot and Northern blot were performed to observe the expression of MDR1 in MCF-7/ADR. Adriamycin accumulation was measured by flow cytometry (FCM). ResultsGSP ( 1.2 mg/L ,2.4 mg/L) inhibited the expression of P-gp to 80.83%( t =5.58,P <0.05) and 50.81%( t =10.96,P <0.01) respectively,compared with the control. With GSP (1.2 mg/L, 2.4 mg/L ),Northern blot analysis showed that the transcription of MDR1 were 60.92% ( t =16.03,P <0.005) and 42.05%( t =21.70,P <0.005) of the control. The cellular accumulation of adriamycin was markedly increased at the presence of 2.4 mg/L GSP. Conclusions GSP has a multidrug resistance reversal action . It may be a potential agent for modulating the multidrug resistance phenotype in cancer cells.