摘要
采用中效原理评价恰玛古多糖(BRCPe)联合顺铂(cisplatin)对人肝癌细胞(HepG2)的协同抑制作用,并考察体外对HepG2凋亡、迁移等生物学行为的影响,阐明其可能的分子机制。用不同浓度的BRCPe、Cisplatin及二者联合用药作用于HepG2细胞24 h、48 h, MTT法检测药物对HepG2细胞的增殖抑制作用,根据中效原理分析两药联合作用的效应(fa)与联用指数(CI)的关系,探讨药物间的相互作用;细胞划痕实验考察药物单独或联用后对细胞迁移的影响;Hoechst 33258荧光染色观察细胞凋亡形态;Western印迹检测凋亡相关蛋白质表达水平。两种药物单独或联用均显著抑制了HepG2细胞的增殖;由中效原理计算得知,两药联用抑制HepG2细胞增殖作用增强,存在协同效应;划痕实验显示,两药联用对HepG2细胞的迁移抑制效果优于对照组和单独作用组;Hoechst 33258荧光染色揭示,联合用药组细胞核染色体凝聚现象加剧;Western印迹分析结果显示,联合用药细胞内Bax, Cyt C,Cleaved Caspase-9/7/3,PARP表达上调,Bcl-2下调。BRCPe与Cisplatin联用对HepG2产生协同抑制效应,其协同机制与诱导线粒体途径介导的细胞凋亡有关。
We aim to evaluate the synergistic inhibition of the Brassica rapa L.polysaccharide(BRCPe)combined with Cisplatin on human hepatocellular carcinoma cells(HepG2)on the intermediate efficacy principle,and to investigate the effects of the combination of BRCPe and cisplatin on the biological behaviors of cell apoptosis and migration in vitro,and to clarify its possible molecular mechanism.HepG2 cells were treated with BRCPe,Cisplatin and their combination at different concentrations for 24 h and 48 h.The MTT assay was employed to detect the inhibitory effect of drugs on the proliferation of HepG2 cells.The relationship between the effect(fa)and the combination index(CI)of the two drugs was analyzed according to the principle of medium-effect,and the interaction between drugs was discussed.The cell scratch test was used to investigate the effects of the drug alone or in combination on cell migration;Hoechst 33258 fluorescent staining was applied to observe the cell morphology of apoptosis;Western blotting was analyzed to detect the expression level of apoptosis-related proteins.Both drug alone or in combination had a significant inhibitory effect on the proliferation of HepG2 cells;the synergistic effect of the combination of two drugs on the proliferation of HepG2 cells was enhanced as calculated by the medium-effect principle;the scratch test showed that the migration inhibition effect of the combination of two drugs on HepG2 cells was better than that of the control group and BRCPe,Cisplatin group alone;Hoechst 33258 revealed that nuclear chromosome condensation was aggravated in the combination group;Western blotting showed that the expression of Bax,Cyt C,Caspase-9/7/3 and PARP were up-regulated and the expression of Bcl-2 was down-regulated in the combination group.BRCPe combined with Cisplatin has a synergistic inhibitory effect on HepG2,and its synergistic mechanism is related to the induction of apoptosis mediated by the mitochondrial pathway.
作者
孔涵蕊
李欣奕
张心慧
陈赛翕
刘政良
努尔阿米乃·麦麦提
李金玉
KONG Han-Rui;LI Xin-Yi;ZHANG Xin-Hui;CHEN Sai-Xi;LIU Zheng-Liang;NURAMINA Ma-Mat;LI Jin-Yu(Xinjiang Key Laboratory of Special Species Conservation and Regulatory Biology,Key Laboratory of Special Environment Biodiversity Application and Regulation in Xinjiang,College of Life Science,Xinjiang Normal University,Urumqi 830054,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2023年第4期605-614,共10页
Chinese Journal of Biochemistry and Molecular Biology
基金
新疆维吾尔自治区自然科学基金项目(No.2021D01A119)
新疆特殊环境物种多样性应用与调控实验室(No.XJTSWZ-2021-02)
新疆维吾尔自治区“百名青年博士引进计划”天池博士(No.BS2019003)项目资助。
关键词
肝癌
恰玛古多糖
顺铂
中效原理
协同效应
hepatocellular carcinoma(HCC)
Brassica rapa L.polysaccharide
cisplatin
medium-effect principle
synergistic effect