摘要
目的研究菊科植物肿柄菊中tagitinin F对人三阴性乳腺癌MDA-MB-231细胞增殖、诱导凋亡的影响及其作用机制。方法MTT法检测细胞活力;incucyte S3长时间活细胞动态成像与分析系统观测细胞生长状况;克隆形成实验检测细胞体外克隆形成能力;流式细胞术检测细胞周期和凋亡;DCFH-DA探针检测细胞内活性氧(ROS)水平;Western blot检测内质网应激相关蛋白的表达水平。结果不同浓度的tagitinin F处理MDA-MB-231细胞后,细胞活力随着其浓度的增加显著降低;并且,tagitinin F能够抑制MDA-MB-231细胞体外克隆形成能力,诱导的细胞形态改变提示细胞发生了凋亡。进一步的流式细胞术检测表明tagitinin F可以诱导细胞周期阻滞在G_(2)/M期并诱导细胞凋亡。Tagitinin F可以使细胞内ROS积累并且增加内质网应激相关蛋白Bip、PDI、Calnexin、Ero1-Lα、IRE1α的表达,并诱导细胞凋亡。结论Tagitinin F在乳腺癌MDA-MB-231细胞中可以通过ROS的累积激活内质网应激反应,并通过诱导细胞凋亡抑制细胞增殖。
Objective To determine the effect of tagitinin F on the growth of human triple negative breast cancer MDA-MB-231 cells and its mechanism.Methods MDA-MB-231 cells were treated with tagitinin F and at various concentrations.The proliferation and morphological changes of cells treated with tagitinin F were checked by the MTT assay and the incucyte S3 long-term live cell dynamic imaging and analysis system,respectively.The colony formation assay was used to detect tumorigenic ability in vitro.The flow cytometry was used to detect the cell cycle and apoptosis.The intracellular reactive oxygen species(ROS)levels were detected by DCFH-DA probe.Western blot was used to evaluate the expression level of endoplasmic reticulum stress-related proteins.Results After the treatment with different concentrations of tagitinin F in breast cancer MDA-MB-231 cells,the cell viability decreased significantly in a dose-dependent manner.The in vitro colony formation ability was inhibited as well.Morphologically,the cells treated with tagitinin F shrank which indicated apoptosis.Further evaluation with flow cytometry confirmed that tagitinin F induced the cell cycle arrest at G_(2)/M phase and apoptosis.Mechanically,accumulation intracellular ROS was observed when the cells were treated with tagitinin F,and tagitinin F treatment increased the expression levels of endoplasmic reticulum stress-related proteins,including Bip,PDI,Calnexin,Ero1-Lα,and IRE1α.Conclusion Tagitinin F can inhibit the cell proliferation and induce the apoptosis in triple negative breast cancer MDA-MB-231 cells.Tagitinin F-induced apoptosis is possibly mediated by ROS accumulation and thereafter activation of endoplasmic reticulum stress.
作者
王娟
杨旭
赵越勤
丁骁
夏海滨
赵逾涵
WANG Juan;YANG Xu;ZHAO Yue-qin;DING Xiao;XIA Hai-bin;ZHAO Yu-han(Key Laboratory of Gene Therapy,School of Life Sciences,Shaanxi Normal University,Xi'an 710119;State Key Laboratory of Phytochemistry and Plant Resources in West China,Kunming Institute of Botany,Chinese Academy of Sciences,Kunming 650201)
出处
《中南药学》
CAS
2021年第5期789-794,共6页
Central South Pharmacy
基金
国家自然科学基金(No.81703393)
云南省自然科学基金面上项目(No.202001AT070055)。
