摘要
目的:研究苦参碱对人乳腺癌MCF-7细胞自噬及凋亡的影响,并分析其机制,探索二者之间潜在的关系。方法:采用MTT法和Annexin V/PI双染法检测细胞增殖活力和凋亡率;自噬双标腺病毒法(mRFP-GFP-LC3)和透射电镜法检测细胞自噬水平;Western Blot检测自噬相关蛋白Beclin-1、LC3及凋亡相关蛋白Caspase-3、cleaved-Caspase-3、cleaved-Caspase-9的表达;运用自噬抑制剂(3-MA)和凋亡抑制剂(Z-VAD-FMK)对上述相关指标进行干预。结果:苦参碱在未加入抑制剂干预的情况下可显著抑制MCF-7细胞的增殖,提高凋亡率和自噬水平,可上调自噬相关蛋白Beclin-1、LC3和凋亡相关蛋白cleaved-Caspase-3、cleaved-Caspase9的表达(P<0.05或P<0.01)。加入自噬抑制剂(3-MA)后,细胞存活率显著降低,凋亡率显著升高(P<0.05);加入凋亡抑制剂(Z-VAD-FMK)后,自噬水平显著升高(P<0.05)。结论:苦参碱可诱导人乳腺癌MCF-7细胞凋亡和细胞自噬,二者之间可能存在着互相调控的关系。
Objective:To study the effect of matrine on autophagy and apoptosis of human breast cancer MCF-7 cells,to analyze the mechanism and explore the potential relationship between them.Methods:MTT assay and Annexin V/PI double staining were used to detect cell proliferation activity and apoptosis rate.Autophagy double labeled adenovirus assay(mRFP-GFP-LC3)and transmission electron microscopy were used to detect the autophagy level.Western Blot was used to detect the expressions of autophagy related proteins Beclin-1,LC3 and apoptosis related proteins Caspase-3,cleaved-Caspase-3,cleaved-Caspase-9.Autophagy inhibitor(3-MA)and apoptosis inhibitor(Z-VAD-FMK)were used to intervene the above related indexes.Results:Matrine could significantly inhibit the proliferation of MCF-7 cells without the intervention of inhibitors,improve the apoptosis rate and autophagy level,up-regulate the expressions of autophagy related proteins Beclin-1,LC3 and apoptosis related proteins cleaved-Caspase-3,cleaved-Caspase-9(P<0.05 or P<0.01).After the addition of autophagy inhibitor(3-MA),the cell survival rate decreased significantly and the apoptosis rate increased significantly(P<0.05).Autophagy level increased significantly after the addition of apoptosis inhibitor(Z-VAD-FMK)(P<0.05).Conclusion:Matrine can induce apoptosis and autophagy of human breast cancer MCF-7 cells,and there may be a mutual regulatory relationship between them.
作者
贾绍华
孙萌遥
丁海鑫
金诗鹏
JIA Shao-hua;SUN Meng-yao;DING Hai-xin;JIN Shi-peng(College of Pharmacy/Pharmaceutical Engineering Technology Research Center,Harbin University of Commerce,Harbin 150076,China)
出处
《中药材》
CAS
北大核心
2023年第3期724-729,共6页
Journal of Chinese Medicinal Materials
基金
黑龙江省自然科学基金重点项目(ZD2018016)
