基于转录组学探讨欧前胡素改善乳腺癌多柔比星耐药的机制  

Mechanism of imperatorin in ameliorating doxorubicin resistance of breast cancer based on transcriptomics

作　　者：李意婷 董伟 梁新丽 王虎 邱雨美 丁小云 张浩 鲍会云 李娴晰 汤喜兰 LI Yiting;DONG Wei;LIANG Xinli;WANG Hu;QIU Yumei;DING Xiaoyun;ZHANG Hao;BAO Huiyun;LI Xianxi;TANG Xilan(School of Pharmacy,Jiangxi Science&Technology Normal University,Nanchang 330013,China;Key Laboratory of Modern Preparation of Chinese Medicine,Ministry of Education,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西科技师范大学药学院,南昌330013 [2]江西中医药大学现代中药制剂教育部重点实验室,南昌330004

出　　处：《中国药房》2025年第5期529-534,共6页China Pharmacy

基　　金：国家自然科学基金项目(No.82060733);江西省卫生健康委科研课题(No.202311141);江西科技师范大学创新训练项目(No.XJ202411318115);江西中医药大学校级课题(No.jzyjg-2023-07)。

摘　　要：目的研究欧前胡素(IMP)对乳腺癌多柔比星(DOX)耐药的逆转作用及可能机制。方法采用MTT法考察IMP最大无毒质量浓度(100μg/mL)与DOX不同质量浓度(12.5、25、50、75、100μg/mL)联用对MCF-7/DOX细胞增殖的影响;将MCF-7/DOX细胞分为空白对照组(1‰二甲基亚砜)、DOX组(50μg/mL)、IMP+DOX组(100μg/mL IMP+50μg/mL DOX)和IMP组(100μg/mL),检测各组细胞中多药耐药蛋白1(MDR1)、多药耐药相关蛋白1的mRNA及其蛋白表达水平;采用转录组测序技术及基因本体(GO)富集分析、京都基因与基因组百科全书(KEGG)通路富集分析筛选IMP改善乳腺癌DOX耐药相关的通路和靶点,并进行验证。结果与单用DOX比较,IMP与DOX联用后,DOX对MCF-7/DOX细胞的半数抑制浓度从81.965μg/mL下降至43.170μg/mL,耐药逆转倍数为1.90;且联用可显著下调MDR1的mRNA表达(P<0.05)。GO富集分析和KEGG通路富集分析结果显示,IMP逆转乳腺癌DOX耐药主要与调节解毒、多生物过程、细胞杀伤等生物过程有关,主要涉及p53信号通路,关键靶点为组成型光形态建成蛋白1(COP1)、细胞周期蛋白E1(CCNE1)、生长停滞和DNA损伤诱导蛋白45A(GADD45A)及GADD45B。验证实验结果显示,与DOX组比较,IMP+DOX组细胞中COP1的mRNA表达有上调趋势,但CCNE1、GADD45A及GADD45B的mRNA表达均显著下调(P<0.05)。结论IMP逆转乳腺癌DOX耐药的作用可能与调控p53信号通路中COP1、CCNE1、GADD45A及GADD45B的表达有关。OBJECTIVE To investigate the ameliorative effect and potential mechanism of imperatorin(IMP)on doxorubicin(DOX)resistance in breast cancer.METHODS The effects of maximum non-toxic concentration(100μg/mL)of IMP combined with different concentrations of DOX(12.5,25,50,75,100μg/mL)on the proliferation of MCF-7/DOX cells were determined by MTT method.MCF-7/DOX cells were divided into blank control group(1‰dimethyl sulfoxide),DOX group(50μg/mL),IMP+DOX group(100μg/mL IMP+50μg/mL DOX)and IMP group(100μg/mL).mRNA and protein expressions of multidrug resistance protein 1(MDR1)and multidrug resistance-associated protein 1 in each group were measured.The relevant pathways and targets involved in the improvement of DOX resistance in breast cancer cells by IMP were screened and validated by using transcriptome sequencing technology,along with gene ontology(GO)enrichment analyses and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.RESULTS Compared with DOX alone,the combination of IMP and DOX reduced the half inhibitory concentration of DOX on MCF-7/DOX cells from 81.965μg/mL to 43.170μg/mL,the reverse fold was 1.90,and the mRNA expression of MDR1 was significantly down-regulated(P<0.05).The results of GO enrichment analyses and KEGG pathway enrichment analyses indicated that the reversal of DOX resistance in breast cancer by IMP was mainly associated with the regulation of biological processes such as detoxification,multiple biological processes,and cell killing.The main pathway involved was the p53 signaling pathway,and the key targets mainly included constitutively photomorphogenic protein 1(COP1),cyclin E1(CCNE1),growth arrest and DNA damage-inducible protein 45A(GADD45A)and GADD45B.The results of the verification experiments showed that compared with DOX group,there was a trend of up-regulation of COP1 mRNA,and significant downregulation of CCNE1,GADD45A,and GADD45B mRNA expression in IMP+DOX group(P<0.05).CONCLUSIONS The effect of IMP in ameliorating DOX resistance in breast cancer is

关 键 词：欧前胡素 乳腺癌 多柔比星耐药 转录组学 p53信号通路 

分 类 号：R965[医药卫生—药理学] R285[医药卫生—药学]