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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。
Objective To study the effects of norcantharidin (NCTD) and evodiamine (EVO) on the proliferation and apoptosis of HepG2 hepatocellular carcinoma cells. Methods Different concentrations of NCTD and different concentrations of EVO were divided into single drug treatment group and NCTD, EVO combined treatment group, another set of non-drug control group, acting on HepG2 cells. MTT assay was used to detect the proliferation of HepG2 cells treated with NCTD and EVO alone. The cell cycle was detected by flow cytometry. The apoptosis of HepG2 cells was detected by annexinⅤ-FITC / PI double staining and flow cytometry. Western blot was used to detect the expression of Bak and Bcl-2 protein. Results MTT assay showed that NCTD and EVO could inhibit the growth of HepG2 cells. The combined treatment of both drugs showed significant synergistic inhibitory effect (CI <1). NCTD and EVO up-regulated G2 / M phase compared with single treatment (P <0.05). The blockage rates of G2 / M phase were EVO (36.13 ± 1.63)%, NCTD (10.67 ± 0.89)% and combined treatment (73.42 ± 1.92)%, respectively. The apoptotic rate was significantly increased from (15.78 ± 3.08)% and NCTD (11.47 ± 1.60)% to (21.86 ± 2.70)% (P <0.05), respectively. Western blot results showed that compared with the single treatment, Bax protein expression increased, Bcl-2 expression decreased (P <0.05). Conclusions The combined treatment of NCTD and EVO can inhibit the proliferation and promote the apoptosis of hepatoma cells.