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

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (5): 512-517.

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

李絮¹，代谊¹，黄家凤¹，温纯洁¹，邹镇¹，吴兰香¹，王果²，周宏灏^{1, 2}

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，长沙 400078，湖南

收稿日期:2016-09-27 修回日期:2016-12-19 出版日期:2017-05-26 发布日期:2017-05-27
通讯作者: 吴兰香，女，博士，副研究员，研究方向：临床药理学。 Tel:15023250939 E-mail: lxwu2008@126.com
作者简介:李絮，女，硕士研究生，研究方向：临床药理学。 Tel:15123826589 E-mail:549290095@qq.com
基金资助:
国家自然科学基金（81473284,81102511）；重庆市自然科学基金（cstc2011jjA10004）；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）

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

LI Xu1, Dai Yi¹, HUANG Jiafeng ¹, WEN Chunjie¹, ZOU Zhen¹, WU Lanxiang ¹, WANG Guo², ZHOU Honghao ^1,2

¹ Institute of Life Science, Chongqing Medical University, Chongqing 400016, China; ²Institute of Clinical Pharmacology, Central South University, Changsha 400078, Hunan, China

Received:2016-09-27 Revised:2016-12-19 Online:2017-05-26 Published:2017-05-27

摘要/Abstract

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；Realtime 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22.03 μmol/L和27.46 μmol/L；10 μmol/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。

关键词: 姜黄素, MCF-7, MCF-7/DOX, 多药耐药基因1, 乙酰化

Abstract:

AIM: To study the effect of curcumin on histone acetylation and the expression level of MDR1 (multidrug resistance protein 1) in breast cancer cells MCF-7 and MCF-7/DOX. METHODS: The effect of curcumin on the growth proliferation 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 expression levels of MDR1 in MCF-7 and MCF-7/DOX cells. The acetylation levels of MDR1 promoter region related histone H3, H4 were detected by chromatin immunoprecipitation assay. RESULTS:Curcumin 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 sensitivity of MCF-7 and MCF-7/DOX cells were significantly increased after the treatment of 10 μmol/L curcumin for 72 h, and the values of IC50 were decreased by 57.14% and 54.52% (P<0.05), respectively. 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), respectively. The MDR1 promoter region related levels of histone H3 and H4 acetylation in MCF-7 and MCF-7/DOX cells were significantly induced by curcumin (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.

Key words: curcumin, MCF-7, MCF-7/DOX, multidrug resistance protein 1, acetylation

中图分类号:

R965.2

李絮，代谊，黄家凤，温纯洁，邹镇，吴兰香，王果，周宏灏. 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[J]. 中国临床药理学与治疗学, 2017, 22(5): 512-517.

LI Xu, Dai Yi, HUANG Jiafeng, WEN Chunjie, ZOU Zhen, Wu Lanxiang, WANG Guo, ZHOU Honghao. Curcumin increases MDR1 expression in breast cancer MCF-7 and MCF-7/DOX cells through regulating histone acetylation[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(5): 512-517.

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