姜黄素对乳腺癌MCF-7与MCF-7/DOX细胞中P-糖蛋白表达的影响及其机制研究  

Effect of curcumin on the P-glycoprotein expression in MCF-7 and MCF-7 / DOX cells and its mechanism

作　　者：王寅声[1] 李晓智[1] 沈杰[2] 李絮[3] 温纯洁[3] 吴兰香[3] 周宏灏[3] 

机构地区：[1]重庆医科大学附属第一医院口腔科,重庆400016 [2]皖南医学院弋矶山医院药物临床评价中心 [3]重庆医科大学生命科学研究院,重庆400016

出　　处：《中国临床药理学与治疗学》2015年第7期769-774,共6页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：重庆市自然科学基金(cstc2011jj A10004);国家自然科学基金(81473284);重庆市医学科研计划项目(2013-2-150);重庆市渝中区科技计划项目(20130127)

摘　　要：目的:研究姜黄素对人乳腺癌MCF-7与MCF-7/DOX细胞中P-糖蛋白表达的影响及其作用机制。方法:MTS比色法检测姜黄素对MCF-7与MCF-7/DOX细胞多柔比星敏感性的影响;实时定量PCR与Western Blot方法检测姜黄素处理后,MCF-7与MCF-7/DOX细胞中P-糖蛋白在mRNA与蛋白质水平的表达变化;流式荧光法检测姜黄素对MCF-7与MCF-7/DOX细胞中多柔比星药物浓度的影响;亚硫酸氢盐处理后测序(bisulfite sequencing PCR,BSP)法检测姜黄素处理前后MCF-7与MCF-7/DOX细胞中P-糖蛋白编码基因多药耐药基因1(multidrug resistance protein1,MDR1)启动子区甲基化状态的改变。结果:与对照组相比,姜黄素处理后,MCF-7和MCF-7/DOX细胞多柔比星IC50值显著下降(P<0.05);姜黄素处理后,MCF-7和MCF-7/DOX细胞中的P-糖蛋白在mRNA及蛋白质水平的表达均有显著上调(P<0.05),而细胞内多柔比星的药物浓度下降;MCF-7和MCF-7/DOX细胞中MDR1基因启动子区域的甲基化程度在姜黄素处理后明显降低(P<0.05)。结论:姜黄素可增强乳腺癌MCF-7与MCF-7/DOX细胞多柔比星敏感性,但同时可显著诱导P-糖蛋白表达水平增高,从而降低细胞内多柔比星药物浓度,此作用与其对MDR1基因去甲基化功能密切相关。AIM： To investigate the effect of curcumin on the expression levels of P-glycoprotein in human breast cancer MCF-7 and MCF-7 / DOX cells and its mechanism. METHODS： The effect of curcumin on the doxorubicin sensitivities of MCF-7and MCF-7 / DOX cells were evaluated by MTS assay. Real-time PCR and Western Blot assays were used to detect the expression levels of P-glycoprotein in MCF-7 and MCF-7 / DOX cells. Intracellular doxorubicin accumulation in cells were measured by flow cytometry（ FCM）. The methylation status in the promoter region of MDR1 gene were analyzed by BSP（ bisulfite sequencing PCR） assay. RESULTS： Compared with control group,the doxorubicin IC50 values of MCF-7 and MCF-7 / DOX cells were decreased after curcumin treatment（ P〈0. 05）. Curcumin treatment significantly increased the expression levels of P-glycoprotein both in mRNA and protein level（ P〈0. 05）,and decreased the concentration of doxorubicin in MCF-7 and MCF-7 / DOX cells. BSP assay showed that the methylation ratios in promoter region of MDR1 gene in MCF-7 and MCF-7 / DOX cells were decreased after curcumin treatment（ P〈0. 05）. CONCLUSION： Curcumin can increase the doxorubicin sensitivities of MCF-7 and MCF-7 / DOX cells,but simultaneously,it can also increase the expression level of P-glycoprotein,which may closely related to its DNA demethylation function.

关 键 词：姜黄素 MCF-7细胞 MCF-7/DOX细胞 P-糖蛋白 DNA甲基化 

分 类 号：R965.2[医药卫生—药理学]