【摘要】 目的研究去甲斑蝥素(NCTD)与吴茱萸碱(EVO)联合处理对肝癌HepG2细胞增殖和凋亡的影响。方法以不同浓度的NCTD和不同浓度的EVO分别组成单药处理组和NCTD、EVO联合处理组,另设不加药的对照组,作用于HepG2细胞。MTT法检测NCTD、EVO单药及联用对HepG2细胞的增殖抑制作用,流式细胞术检测细胞周期,annexinⅤ-FITC/PI双染色结合流式细胞术检测药物对HepG2细胞凋亡的影响。Western blot法检测Bak、Bcl-2蛋白的表达。结果 MTT法证明NCTD和EVO对HepG2细胞的生长有抑制效果,两药联合处理具有明显的协同抑制作用(合用指数CI<1);与单一处理组相比,NCTD联合EVO上调G2/M期阻滞率(P<0.05),G2/M期阻滞率分别为EVO(36.13±1.63)%、NCTD(10.67±0.89)%、联合处理(73.42±1.92)%;此外,两药联合处理的肝癌细胞凋亡率由EVO(15.78±3.08)%、NCTD(11.47±1.60)%,显著上升到联合处理的(21.86±2.70)%(P<0.05);Western blot结果显示,与单独处理相比较,联合处理组Bax蛋白表达水平增高,Bcl-2表达降低(P<0.05)。结论 NCTD和EVO联合处理抑制肝癌细胞增殖并促进细胞凋亡。更多还原

【Abstract】 Objective To investigate the combined effect of evodiamine( EVO) and norcantharidin( NCTD) on HepG2cells. Methods HepG2 cells were treated with EVO or /and NCTD at different concentrations in vitro,with blank culture medium as the negative control. The inhibitory effect of EVO or /and NCTD was determined by MTT assay. Cell cycle and cell apoptosis were assessed by flow cytometry( FCM) combined with PI staining and annexin V-FITC /PI,respectively. The expressions of Bax and Bcl-2 were examined by Western blotting. Results MTT assay showed that NCTD and EVO had restraining effect on the growth of HepG2 cells. The combination of NCTD and EVO had significant synergistic inhibition effect( CI <1). The arrest rate of G2/M phase was( 36.13 ±1.63) % in the EVO induced group and( 10.67 ±0.89) % in the NCTD induced group,and the combination of EVO and NCTD up-regulated the rate to( 73. 42 ± 1. 92) %( P < 0. 05). In addition,the apoptosis rate of HepG2 cells treated by two-drug combination increased significantly from( 15. 78 ± 3. 08) % in the EVO group and( 11. 47 ± 1. 60) % in the NCTD group to( 21. 86 ± 2. 70) % in the EVO combined with NCTD group( P < 0. 05).Western blotting demonstrated that the expression level of Bax protein went up significantly( P < 0. 05),meanwhile Bcl-2protein dropped significantly( P < 0. 05) in the combined induced group. Conclusion EVO combined with NCTD showed synergetic effect on anti-proliferation and pro-apoptosis in HepG2 cells.更多还原