桑树类黄酮对三阴性乳腺癌细胞MDA-MB231的细胞毒性作用初探
Study on the Cytotoxic Effects of Mulberry Flavonoids on Breast Cancer Cells
钱鹏 1袁刚祥 1何宁佳1
作者信息
- 1. 资源昆虫高效养殖与利用全国重点实验室(西南大学),重庆 400715
- 折叠
摘要
桑树中富含的类黄酮具有多种药理活性.采用 3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲酯基)-2-(4-磺苯基)-2 H-四唑试剂(MTS)染色和实时荧光定量 PCR(RT-qPCR)等方法,就桑树中类黄酮对乳腺癌细胞 MDA-MB231 的细胞毒性作用进行初步探索.实验结果显示,供试的 4 种桑树类黄酮中,戊烯基化类黄酮亚类对乳腺癌细胞的毒性作用强于非戊烯基化类黄酮.其中,戊烯基化类黄酮——桑根酮 C 整体呈现出更强的细胞毒性作用,当桑根酮C的浓度达到 20 μmol/L时能显著抑制 MDA-MB231 增殖.胸腺嘧啶脱氧核苷类似物(EDU)检测细胞增殖等实验结果显示,20 μmol/L桑根酮C处理 MDA-MB231 后,细胞中 DNA的复制明显减少,体外克隆能力显著减弱;RT-qPCR检测细胞周期阻滞和DNA损伤相关的周期蛋白依赖性激酶抑制剂 1A 基因(CDKN1A)和生长阻滞与DNA损伤基因(GADD45A)的转录水平显著上升,而 DNA复制相关 S期激酶相关蛋白 2 基因(SKP2)和增殖细胞核抗原基因(PCNA)的转录水平则显著降低.此外,用桑根酮 C 处理 MDA-MB231 后,出现细胞形态变圆变亮和变小、细胞中线粒体膜电位丢失和 ROS积累等现象,以及促凋亡基因 phorbol-12-myristate-13-acetate 诱导蛋白 1 基因(PMAIP1)的转录水平显著上升,抑制凋亡基因Bcl2 的转录水平显著下降,提示桑根酮C可能诱导细胞发生凋亡.
Abstract
The flavonoids abundant in mulberry have various pharmacological activities.In this study,the cytotoxic effects of mulberry flavonoids on breast cancer cells MDA-MB231 were preliminarily explored using methods such as 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium(MTS)assay and real-time quantitative polymerase chain reaction(RT-qPCR).The experi-mental results showed that among the four types of mulberry flavonoids tested,the prenylated flavonoid subfamily exhibited stronger cytotoxic effects on breast cancer cells than the non-prenylated flavonoids.A-mong them,the prenylated flavonoid Sanggenon C showed the strongest overall cytotoxic effect.When the concentration of Sanggenon C reached 20 μmol/L,it significantly inhibited the proliferation of MDA-MB231 cells.Experiments such as 5-ethynyl-2'-deoxyuridine(EDU)cell proliferation assay showed that after treatment with 20μmol/L Sanggenon C,DNA replication in MDA-MB231 cells was significantly re-duced,and the in vitro cloning ability was significantly weakened.RT-qPCR analysis showed that the tran-scription levels of Cyclin-Dependent Kinase Inhibitor 1 A(CDKN1A)and Growth Arrest and DNA Dam-age-inducible gene 45 A(GADD45A),genes related to cell cycle arrest and DNA damage,were signifi-cantly increased,while the transcription levels of S-phase kinase-associated protein 2(SKP2)and Prolifer-ating Cell Nuclear Antigen(PCNA),genes related to DNA replication,were significantly decreased.Ad-ditionally,after treatment with Sanggenon C,the cells changed their morphology to become rounder and brighter,with reduced cell volume.Loss of mitochondrial membrane potential and accumulation of ROS suggested that Sanggenon C may induce apoptosis in the cells.The transcription level of the pro-apoptotic gene Phorbol-12-myristate-13-acetate-induced protein 1(PMAI P1)was significantly increased,while the transcription level of the anti-apoptotic gene Bcl2 was significantly decreased,further indicating that Sanggenon C may induce apoptosis in the cells.
关键词
桑树/类黄酮/细胞毒性/凋亡/增殖抑制Key words
Morus L./Flavonoids/Cytotoxicity/Apoptosis/Proliferation inhibition引用本文复制引用
基金项目
国家重点研发计划(2022YFD1201602)
西南大学创新研究先导计划(2035)(SWU-XDZD22008)
重庆市基础研究与前沿技术项目(cstc2021yszxjcyj0004)
出版年
2024
NETL
NSTL
万方数据