摘要
目的:探讨薯蓣皂苷(dioscin,DIO)对人乳腺癌阿霉素(doxorubicin,DOX)耐药的影响及其可能的作用机制。方法:采用MTT法检测DIO不同浓度对MCF-7/DOX细胞活力的影响,并运用MTT、Western blot和qRT-PCR等方法考察DIO对MCF-7/DOX细胞DOX耐药的逆转作用。采用网络药理学预测DIO干预乳腺癌DOX耐药的潜在靶点,利用STRING数据库和Cytoscape 3.9.1软件构建PPI网络,筛选核心靶点,通过DAVID数据库进行GO功能富集分析和KEGG通路富集分析。最后,采用qRT-PCR实验对核心靶点进行验证。结果:DIO质量浓度在0.375~1.56μg·mL^(–1)时对MCF-7/DOX细胞活力无明显影响。0.75μg·mL^(–1)DIO与不同浓度DOX联用后,DOX对MCF-7/DOX细胞的IC50下降,DIO的逆转耐药倍数为2.02,且MCF-7/DOX细胞中MDR1、MRP1蛋白和mRNA表达下调(P<0.05),表明DIO增强了MCF-7/DOX细胞对DOX的敏感性,逆转MCF-7/DOX细胞耐药。网络药理学分析显示DIO逆转乳腺癌阿霉素耐药的潜在靶点78个,其中核心靶点包括AKT1、TP53、BCL2、IL-6、CASP3、JUN等,主要涉及脂质和动脉粥样硬化、癌症等信号通路。验证实验结果显示0.75μg·mL^(–1)DIO与DOX联用后,上调TP53和CASP3 mRNA表达(与对照组比较,P<0.05),下调AKT1和IL-6 mRNA表达(与DOX比较,P<0.05),有下调BCL2 mRNA表达的趋势,对JUN mRNA无明显影响。结论:DIO可通过作用于AKT1、TP53、BCL2、IL-6、CASP3等核心靶点而逆转乳腺癌DOX耐药。
OBJECTIVE To explore the effect of dioscin(DIO)on doxorubicin(DOX)resistance in human breast cancer MCF-7/DOX cell and elucidate its possible mechanism.METHODS MTT assay was utilized for detecting the viability of MCF-7/DOX cell after treating with different concentrations of DIO.MCF-7/DOX cell was exposed to non-toxic concentrations of DIO plus DOX for observing the reversal effect of DIO on DOX resistance by methylthiazolyldiphenyl tetrazolium(MTT),West⁃ern blot and quantitative real-time polymerase chain reaction(qRT-PCR).Network pharmacology was utilized for predicting the potential targets of DIO against DOX resistance in breast cancer.Protein-protein interaction(PPI)network was constructed through a STRING database and Cytoscape 3.9.1 software for selecting core targets.Gene Ontology(GO)and Kyoto Encyclo⁃pedia of Genes and Genomes(KEGG)pathway enrichment analyses were further performed through the David database.Finally qRT-PCR was performed to verify the mRNA expression of core targets.RESULTS At a concentration of 0.375−1.56μg·mL^(–1),DIO displayed little cellular cytotoxicity.After exposure to DIO 0.75μg·mL^(–1) plus different concentrations of DOX,half maxi⁃mal inhibitory concentration(IC50)of DOX declined with a reversing multitude of 2.02 and protein/mRNA expressions of MDR1/MRP1 became down-regulated(P<0.05).It implied that DIO reversed DOX resistance.Network pharmacology prediction yielded 78 potential targets for DIO reversing DOX resistance in breast cancer.Some core targets included protein kinase B(AKT1),tumor protein P53(TP53),apoptosis regulator BCL2(BCL2),interleukin-6(IL-6),caspase 3(CASP3)and jun proto-oncogene(JUN)involved in lipid and atherosclerosis,neoplastic pathways and other signal pathways.0.75μg·mL^(–1)DIO plus DOX up-regulated mRNA expressions of TP53 and CASP3(vs.control group,P<0.05)and down-regulated mRNA expressions of AKT1 and IL-6(vs.DOX group,P<0.05).With a tendency of down-regulating BCL2 mRNA,it had no obvi⁃ous effect on JUN mRNA level.CONCLUSION DIO
作者
丁小云
张浩
谢梦蝶
邱雨美
王虎
李停停
董伟
汤喜兰
DING Xiaoyun;ZHANG Hao;XIE Mengdie;QIU Yumei;WANG Hu;LI Tingting;DONG Wei;TANG Xilan(School of Pharmacy,Jiangxi Science&Technology Normal University,Jiangxi Nanchang 330013,China;Key Laboratory of Modern Preparation of Traditional Chinese Medicine,Ministry of Education,Jiangxi University of Chinese Medicine,Jiangxi Nanchang 330004,China;Jiangxi Provincial Key Laboratory of Drug Designs and Evalua-tions,Jiangxi Nanchang 330013,China)
出处
《中国医院药学杂志》
CAS
北大核心
2024年第16期1868-1874,共7页
Chinese Journal of Hospital Pharmacy
基金
国家自然科学基金项目(编号:82060733,81960732)
江西省自然科学基金项目(编号:20224BAB206111)。
关键词
薯蓣皂苷
乳腺癌
阿霉素耐药
网络药理学
dioscin
breast cancer
doxorubicin resistance
network pharmacology
