摘要
目的探讨香贝散体外抗乳腺癌作用及其分子机制。方法体外培养人乳腺癌细胞MCF-7、MDA-MB-231,给予香贝散1、2、3、4、5 mg·mL^(-1)加药处理,对照组不加药。利用MTT法和克隆形成实验检测细胞增殖,利用流式细胞术检测细胞凋亡,细胞划痕和细胞侵袭实验检测细胞迁移和侵袭能力,借助吖啶橙染色法观察细胞自噬,借助转录组测序技术探索香贝散诱导乳腺癌细胞死亡的作用机制,并采用Western blotting法检测AMP依赖的蛋白激酶(AMPK)、哺乳动物雷帕霉素靶蛋白(m TOR)、LC-3、Beclin-1蛋白表达水平。结果与对照组比较,香贝散显著抑制人乳腺癌MCF-7、MDA-MB-231细胞活力(P<0.05、0.001),48 h半数抑制浓度(IC50)分别为2.344、1.961 mg·mL^(-1),且具有时间、浓度相关性;显著抑制MCF-7、MDA-MB-231细胞的集落形成能力;显著诱导MCF-7、MDA-MB-231细胞凋亡(P<0.001);明显抑制人乳腺癌MCF-7、MDA-MB-231细胞的体外迁移能力和侵袭能力;吖啶橙染色实验结果提示香贝散能够诱导MCF-7、MDA-MB-231细胞发生自噬,Western blotting实验结果显示香贝散能够显著上调MCF-7、MDA-MB-231细胞中自噬关键蛋白Beclin-1和LC3-Ⅱ的表达(P<0.01、0.001);转录组测序技术发现差异基因变化最明显的通路包括mTOR信号通路、自噬信号通路等,Western blotting实验结果显示香贝散显著下调mTOR的磷酸化水平(P<0.001),显著上调AMPK的磷酸化水平(P<0.001)。结论香贝散能够显著抑制人乳腺癌细胞的增殖、迁移、侵袭能力,能够诱导凋亡和自噬的发生,激活AMPK/mTOR信号通路可能是其重要机制。
Objective To investigate the in vitro anti-breast cancer effects of the Chinese herbal prescription Xiangbeisan and its molecular mechanisms.Results Human breast cancer MCF-7 and MDA-MB-231 cells were treated with Xiangbeisan 1,2,3,4,or 5 mg:mL-l,with a control group not receiving any drug.MTT assay and clone formation assay were used to detect the effects of Xiangbeisan on the proliferation of human breast cancer MCF-7 and MDA-MB-231 cells,and flow cytometry was used to detect the changes in apoptosis of human breast cancer MCF-7 and MDA-MB-231 cells after the administration of Xiangbeisan.Subsequently,the effects of Xiangbeisan on the migration and invasion ability of human breast cancer MCF-7 and MDA-MB-231 cells were detected using cell scratch and cell invasion assays.Through acridine orange staining,the effect of Xiangbeisan on autophagy in human breast cancer cells was explored.Finally,the transcriptome sequencing technique was used to explore the mechanism of Xiangbeisan-induced breast cancer cell death,and the western blotting experiment was used to detect the protein levels of AMPK,mTOR,LC3,and Beclin-1.Results Compared with the control group,Xiangbeisan significantly inhibited the viability of MCF-7 and MDA-MB-231 cells(P<0.05,0.001),and the median inhibitory concentration(IC_(50))at 48 h were 2.344 and 1.961 mg·mL^(-1),respectively,with time and concentration correlation.The colony formation ability of MCF-7 and MDA-MB-231 cells was significantly inhibited.Apoptosis of MCF-7 and MDA-MB-231 cells was significantly induced(P<0.001).Xiangbeisan obviously inhibited the migration and invasion ability of MCF-7 and MDA-MB-231 cells in vitro.The results of acridine orange staining indicated that Xiangbeisan induced autophagy in MCF-7 and MDA-MB-231 cells.Western blotting experiments showed that Xiangbeisan significantly up-regulated the expressions of Beclin-1 and LC3-II,the key autophagy proteins,in MCF-7 and MDAMB-231 cells(P<0.01,0.001).Transcriptome sequencing assay revealed that the most significant pat
作者
谢锦欣
王文艺
全晓敏
邢昊晴
贾智
王飞
魏雪娇
谭鹏
王柱国
胡仲冬
安超
XIE Jinxin;WANG Wenyi;QUAN Xiaomin;XING Haoqing;JIA Zhi;WANG Fei;WEI Xuejiao;TAN Peng;WANG Zhuguo;HU Zhongdong;AN Chao(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China;Modern Research Center for Traditional Chinese Medicine,Beijing Institute of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;Department of Oncology,Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China)
出处
《药物评价研究》
CAS
北大核心
2024年第6期1249-1258,共10页
Drug Evaluation Research
基金
中央高校基本科研业务费专项资金项目(2023-JYB-JBQN-051)
北京中医药大学国家级人才精准培育计划项目(JZPY202206)
教育部双一流引导专项经费-国际合作平台项目(90020361020006)。
关键词
香贝散
乳腺癌
增殖
迁移
侵袭
凋亡
自噬
AMPK/mTOR信号通路
Xiangbeisan
breast cancer
proliferation
migration
invasion
apoptosis
autophagy
AMPK/mTOR signaling pathway
