姜黄素通过调控组蛋白乙酰化修饰上调乳腺癌MCF-7和MCF-7/DOX细胞中MDR1表达  

Curcumin increases MDR1 expression in breast cancer MCF-7 and MCF-7/DOX cells through regulating histone acetylation

作　　者：李絮[1] 代谊 黄家凤 温纯洁[1] 邹镇[1] 吴兰香[1] 王果[2] 周宏灏[1,2] 

机构地区：[1]重庆医科大学生命科学研究院,重庆400016 [2]中南大学临床药理研究所

出　　处：《中国临床药理学与治疗学》2017年第5期512-517,共6页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金(81473284;81102511);重庆市自然科学基金(cstc2011jj A10004);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(0130127)

摘　　要：目的:从组蛋白乙酰化修饰角度探讨姜黄素上调乳腺癌MCF-7和MCF-7/DOX细胞中多药耐药基因1(multidrug resistance protein 1,MDR1)表达水平的作用机制。方法:CCK-8(Cell Counting Kit-8)法观察姜黄素对MCF-7和MCF-7/DOX细胞生长增殖以及多柔比星敏感性的影响;Realtime PCR和Western blot方法检测姜黄素处理前后两种细胞中MDR1 mRNA和蛋白质表达水平的变化;染色质免疫共沉淀技术(chromatin immunoprecipitation assay,CHIP)检测姜黄素对两种细胞中MDR1启动子区相关组蛋白H3、H4乙酰化水平的影响。结果:不同浓度姜黄素处理72 h对MCF-7和MCF-7/DOX细胞的生长增殖有显著抑制作用,其IC50值分别为22.03μmol/L和27.46μmol/L;10μmol/L姜黄素处理72 h可显著提高两种细胞多柔比星敏感性,多柔比星IC50值分别下降了57.14%和54.52%(P<0.05);姜黄素处理MCF-7和MCF-7/DOX细胞后,MDR1在mRNA水平分别上调(32.90±0.96)和(5.70±0.55)倍(P<0.05),在蛋白质水平分别上调(2.26±0.18)与(2.90±0.21)倍(P<0.05);CHIP结果显示姜黄素对两种细胞中MDR1启动子区相关组蛋白H3、H4的乙酰化水平均有显著上调作用。结论:姜黄素在增强MCF-7和MCF-7/DOX细胞多柔比星敏感性的同时,可通过调控MDR1启动子区相关组蛋白H3、H4的乙酰化水平而上调MDR1的表达。AIM： To study the effect of curcu- min on histone acetylation and the expression level of MDR1 （ multidrug resistance protein 1 ） in breast cancer cells MCF-7 and MCF-7/DOX. METH- ODS ： The effect of curcumin on the growth prolifer- ation and sensitivities of doxorubicin against MCF-7 and MCF-7/DOX cells were evaluated by CCK-8 （Cell Counting Kit-8 ） assay. Real-time PCR and Western blot assays were used to detect the expres- sion levels of MDR1 in MCF-7 and MCF-7/DOX cells. The acetylation levels of MDR1 promoter re- gion related histone H3, H4 were detected by chro- matin immunoprecipitation assay. RESULTS： Cur- cumin inhibited the growth of MCF-7 and MCF-7/ DOX cells with the IC50 values of 22.03 μmoL/L and 27.46 μmol/L, respectively. The doxorubicin sen- sitivity of MCF-7 and MCF-7/DOX cells were signif- icantly increased after the treatment of 10 μmol/L curcumin for 72 h, and the values of IC50 were de creased by 57.14% and 54.52% （P 〈 0. 05 ） , re- spectively. After curcumin treatment, MDR1 mRNA levels were （32.90±0.96） and （5.70±0.55） folds increased in MCF-7 and MCF-7/DOX cells （P 〈 0.05 ） , respectively ; MDR1 protein levels were （2.26 ± 0.18 ） and （2.90 ±0.21 ） folds increased in MCF-7 and MCF-7/DOX cells （P 〈 0.05 ）, re- spectively. The MDR1 promoter region related levels of histone H3 and H4 acetylation in MCF-7 and MCF-7/DOX cells were significantly induced by cur- cumin （P 〈 0.05 ）. CONCLUSION： Curcumin enhances the doxorubicin sensitivities in MCF-7 and MCF-7/DOX cells. Moreover, it induces the MDR1 expression via increasing the MDR1 promoter region related levels of histone H3 and H4 acetylation.

关 键 词：姜黄素 MCF-7 MCF-7/DOX 多药耐药基因1 乙酰化 

分 类 号：R965.2[医药卫生—药理学]