小檗碱和XAV939联合应用可抑制骨肉瘤细胞MG-63的迁移与凋亡OA北大核心CSTPCD
Combined application of berberine and XAV939 inhibits the migration and apoptosis of osteosarcoma cells MG-63
目的:探讨小檗碱(BBR)联合XAV939对骨肉瘤MG-63细胞迁移与凋亡的影响及其可能的机制.方法:培养MG-63细胞,分别加入20～120 μmol/L的BBR和5～60 μmol/L的XAV939,通过CCK-8法测定BBR和XAV939的细胞毒性,采用Chou-Talalay分析法计算两药的联合指数,确定后续实验的干预剂量;将MG-63细胞随机分为空白对照(NC)组、BBR组、XAV939组和BBR+XAV939联合组,采用划痕愈合实验、Transwell小室法检测BBR与XAV939单独或联合处理对MG-63细胞迁移能力的影响,Hoechst 33258染色、JC-1染色及Annexin Ⅴ-FITC/PI双染流式细胞术检测对细胞线粒体膜电位和凋亡的影响,免疫荧光法检测BAX蛋白的表达,WB法检测对细胞中MMP-2表达的影响,qPCR法检测对MMP-2基因表达的影响.结果:BBR和XAV939以剂量依赖方式抑制MG-63细胞的增殖,选定30 μmol/L BBR、7.5 μmol/L XAV939用于后续实验.与NC组相比,BBR(30 μmol/L)、XAV939(7.5 μmol/L)及BBR+XAV939联合处理细胞24 h后,MG-63细胞迁移能力显著降低、凋亡细胞显著增加(均P<0.05),线粒体膜电位下降(P<0.01),MMP-2的基因表达和MMP-2、BAX蛋白水平均降低(均P<0.05),并且,联合组的效应明显强于单药处理且(P<0.05或P<0.01).结论:BBR和XAV939单独或联合应用可以抑制骨肉瘤MG-63细胞的迁移、促进凋亡,其机制可能与迁移相关蛋白MMP-2的表达降低,凋亡相关蛋白BAX的水平增加有关.
Objective:To investigate the effects of berberine(BBR)combined with XAV939 on the migration and apoptosis of osteosarcoma MG-63 cells and its possible mechanisms.Methods:MG-63 cells were cultured,and 20～120 μmol/L of BBR and 5～60 μmol/L of XAV939 were added,respectively.The cytotoxicity of BBR and XAV939 was determined by CCK-8 assay,and the combined index of the two was clarified using Chou-Talalay analysis to determine the intervention dose for the subsequent experiments.MG-63 cells were randomly divided into the blank control(NC)group,the BBR group,the XAV939 group,and the BBR+XAV939 combined group.The effects of BBR and XAV939 treatment alone or in combination on the migratory ability of MG-63 cells were detected by the scratch healing assay and the Transwell assay.Hoechst 33258 staining,JC-1 staining and Annexin Ⅴ-FITC/PI double staining flow cytometry were used to detect the effects on cell mitochondrial membrane potential and apoptosis.Immunofluorescence was used to datect the expression of BAX protein,WB assay was used to detect the effects on the expression of MMP-2 and BAX in the cells,and qPCR was used to detect the effects on the expression of the MMP-2 gene.Results:BBR and XAV939 inhibited the proliferation of MG-63 cells in a dose-dependent manner,and 30 μmol/L BBR,7.5 μmol/L XAV939 were selected for subsequent experiments.Compared with the NC group,24 h after the cells were treated with BBR(30 μmol/L),XAV939(7.5 μmol/L)and BBR+XAV939 combination,the migration ability of MG-63 cells was significantly reduced;apoptotic cells were significantly increased(all P<0.05);the mitochondrial membrane potential was decreased(P<0.01);the gene expression of MMP-2,and MMP-2,BAX protein levels were all reduced(all P<0.05).In addition,the effect of the combination group was significantly stronger than that of the monotherapy group(P<0.05 or P<0.01).Conclusion:BBR and XAV939 alone or in combination can inhibit the migration and promote the apoptosis of MG-63 cells.The mechanism may be related to the decreased expression of migration-related protein MMP-2 and increased level of apoptosis-related protein BAX.
吴昊越;康兵;田志敏;张浩强;李旭升
宁夏医科大学第一临床医学院,宁夏 银川 750004||联勤保障部队第九四〇医院 关节外科,甘肃 兰州 730050联勤保障部队第九四〇医院 关节外科,甘肃 兰州 730050
临床医学
小檗碱XAV939骨肉瘤MG-63细胞迁移凋亡
berberine(BBR)XAV939osteosarcomaMG63 cellmigrationapoptosis
《中国肿瘤生物治疗杂志》 2024 (005)
469-476 / 8
甘肃省青年科技基金(No.20JR5RA588)
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