药学学报, 2018, 53(1): 84-89
引用本文:
祝丽欣, 唐超园, 俞建东, 陈芝, 熊阳. 薏苡仁脂干预MCF-7/DOX细胞中荧光药物动力学及部分耐药蛋白表达[J]. 药学学报, 2018, 53(1): 84-89.
ZHU Li-xin, TANG Chao-yuan, YU Jian-dong, CHEN Zhi, XIONG Yang. Impact of Coix seed oil on fluorescence excretion pharmacokinetics and protein expression in doxorubicin-resistant cells MCF-7/DOX[J]. Acta Pharmaceutica Sinica, 2018, 53(1): 84-89.

薏苡仁脂干预MCF-7/DOX细胞中荧光药物动力学及部分耐药蛋白表达
祝丽欣1,2, 唐超园1, 俞建东1, 陈芝1, 熊阳1
1. 浙江中医药大学药学院, 浙江 杭州 310053;
2. 浙江省食品药品检验研究院, 浙江 杭州 310052
摘要:
本文从薏苡仁脂干预耐药乳腺癌细胞的荧光药物动力学以及对耐药基因和蛋白表达量的影响两方面着手,研究薏苡仁脂逆转乳腺癌细胞耐药的机制。首先通过建立稳定过表达荧光素酶的人乳腺癌耐药细胞株MCF-7/DOXFluc,应用生物发光成像技术(bioluminescence imaging,BLI),实时监测细胞内ATP结合盒(ATP-binding cassette,ABC)转运蛋白底物D-荧光素钾的外排动力学过程,检测薏苡仁脂干预前后D-荧光素钾的胞内动力学变化。结果表明,与空白组相比,薏苡仁脂明显减少MCF-7/DOXFlucD-荧光素钾的外排,增加胞内累积量。此外,采用实时定量基因扩增荧光检测系统和蛋白免疫印迹技术,研究薏苡仁脂干预后MCF-7/DOXFluc中ABC转运子的基因及蛋白表达的变化。结果显示,薏苡仁脂处理后可下调MCF-7/DOXFluc细胞中P-糖蛋白(P-gp/ABCB1)、多药耐药相关蛋白1(MRP1/ABCC1)以及乳腺癌耐药蛋白(BCRP/ABCG2)等耐药基因和蛋白的表达。最后将薏苡仁脂联合多柔比星对薏苡仁脂逆转乳腺癌耐药效果进行体外评价,结果证明薏苡仁脂可增强多柔比星抑制MCF-7/DOX细胞增殖的作用,其最佳联合比例为薏苡仁脂:多柔比星=25:1。薏苡仁脂可以通过抑制ABC转运蛋白的外排功能和下调ABC转运蛋白在肿瘤细胞表达水平,双管齐下逆转化疗药物多柔比星的肿瘤多药耐药。
关键词:   
Impact of Coix seed oil on fluorescence excretion pharmacokinetics and protein expression in doxorubicin-resistant cells MCF-7/DOX
ZHU Li-xin1,2, TANG Chao-yuan1, YU Jian-dong1, CHEN Zhi1, XIONG Yang1
1. College of Pharmacy, Zhejiang Chinese Medical University, Hangzhou 310053, China;
2. Zhejiang Institute for Food and Drug Control, Hangzhou 310052, China
Abstract:
This study was designed to explore the mechanism of Coix seed oil (Coix) impact on the drug resistance, bioluminescence imaging (BLI) and the efflux of D-luciferin potassium salt, the substrate of ABC transporters, in doxorubicin-resistant breast cancer cells. Multidrug resistance (MDR) gene and protein expression were analyzed in the cells by q-PCR and Western blot. First, in order to investigate the effect of the efflux function by ABC protein, a cell line with overexpressed luciferase was established in MCF-7 cell line. BLI was used to monitor the efflux kinetics of D-luciferin potassium salt before and after Coix treament. The results showed that the efflux of D-fluorescein potassium from MCF-7/DOXFluc was lessened when pretreated with Coix, which means that Coix may decrease the efflux of other chemotherapies using ABC transporters. Both of the results of q-PCR and Western blot showed that gene and protein expression of ABC transporters such as ABCG2, ABCC1 and ABCB1 were down-regulated by Coix treatment. The efficacy of Coix reversing MDR was verified with the chemotherapy medication doxorubicin (DOX). MTT assay showed that Coix increased the inhibitory effect of DOX on proliferation of MCF-7/DOX, and the optimal combination of ratio was 25 times that of DOX. The results suggest that Coix may reverse MDR of the substrate of ABC transporters from two aspects, one is to cut down the ABC protein efflux function, and the other is to decrease the quantity of ABC gene and protein expression.
Key words:   
收稿日期: 2017-08-11
DOI: 10.16438/j.0513-4870.2017-0782
基金项目: 国家自然科学基金资助项目(81473434,81774011,81673607).
通讯作者: 熊阳,Tel:86-571-61768158,E-mail:xyxnb@126.com
Email: xyxnb@126.com
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