刘骁 1刘峰 2李鹏运 2郑志兵 2王莹 2肖智勇 1周文霞2
作者信息
- 1. 南京中医药大学,江苏 南京 210023||军事医学研究院,北京 100850||国家安全特需药品全国重点实验室,北京 100039
- 2. 军事医学研究院,北京 100850||国家安全特需药品全国重点实验室,北京 100039
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摘要
Abstract
OBJECTIVE To investigate the in vitro anti-proliferative and anti-metastatic effects of the rimonabant's derivative BSH-5-34 on breast cancer and to explore the underlying mechanism of action.METHODS Human breast cancer cell lines MCF-7,ZR-75-1,and MDA-MB-231 were categorized into three groups:the solvent control group,rimonabant group(1,5 and 10 μmol·L-1),and BSH-5-34 group(1,5 and 10 μmol·L-1).Surface plasmon resonance(SPR)technology was employed to detect the interac-tions between rimonabant and BSH-5-34 on one hand and cannabinoid receptor type 1(CB1).The CCK-8 assay was conducted to assess cell viability while the Transwell assay was used to detect on the other hand the migration of cells.Additionally,an in vitro 3D spheroid proliferation assay was performed to evaluate spheroid formation.Western blotting was employed to assess the expression levels of extracellular signal-regulated kinase(ERK),phosphorylated extracellular signal-regulated kinase(p-ERK),Snail,fibronectin,N-cadherin,E-cadherin,and vimentin in ZR-75-1 cells.The key targets of rimonabant and BSH-5-34 were identified using SwissTargetPrediction,followed by gene ontology(GO)functional analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.RESULTS The equilibrium dissociation constant(KD)of rimonabant was 7.71E-5 M for the CB1 receptor,while the derivative BSH-5-34 showed no detectable affinity(No KD).BSH-5-34 exerted a markedly stronger inhibitory effect than rimonabant at concentrations of 5 and 10 μmol·L-1(P<0.01).Rimonabant(10 μmol·L-1)reduced the relative migration capacity of MDA-MB-231 cells,but had little effect on ZR-75-1 cells.In contrast,BSH-5-34 at both 5 and 10 μmol·L-1 concentrations effectively suppressed the relative migration of both ZR-75-1 and MDA-MB-231 cells(P<0.05).Moreover,BSH-5-34 delivered a more potent inhibitory effect on cell migration than rimonabant(P<0.01).Both compounds inhibited spheroid formation compared with to the solvent control(P<0.05,P<0.01).However,at concentrations of 1,5 and 10 μmol·L-1 of rimonabant,a large number of tumor spheroids were aggre-gated.In contrast,BSH-5-34 almost completely inhibited tumor spheroid formation even at the lowest concentration of 1 μmol·L-1.BSH-5-34 showed notable biological activity.Specifically,BSH-5-34(5 and 10 μmol·L-1)significantly suppressed ERK phosphorylation(P<0.01),and significantly downregulated the expressions of N-cadherin,Snail,fibronectin,and vimentin at 10 μmol·L-1(P<0.05).GO and KEGG enrichment analysis suggested that rimonabant could potentially exert its anti-breast cancer ef-fects through pathways related to immune regulation,vascular remodeling,and platinum resistance.While BSH-5-34 seemed to inhibit breast cancer progression by regulating kinase activity,modulating immu-nometabolic remodeling,and reversing drug resistance.CONCLUSION After structural modification,BSH-5-34 has a more significant inhibitory effect on the proliferation,migration,and tumor spheroid formation of breast cancer cells than the control drug rimonabant.关键词
利莫那班/BSH-5-34/乳腺癌/细胞活力/细胞迁移/体外三维肿瘤模型/上皮内间质转化Key words
rimonabant/BSH-5-34/breast cancer/cell viability/cell migration/in vitro three-dimen-sional tumor model/epithelial-mesenchymal transition分类
医药卫生
