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目的探讨沙利度胺联合表阿霉素抗人肝癌细胞HepG2体外增殖的影响,为肝癌治疗提供依据。方法采用人肝癌细胞株HepG2作为研究对象,共设4组:空白对照组、表阿霉素组(1.0 mg/L)、沙利度胺梯度组(50μg/ml、100μg/ml、200μg/ml和400μg/ml)和联合组(沙利度胺200μg/ml+表阿霉素1.0 mg/L),观察4组用药方案12h、24h、48h和72h时对HepG2细胞的影响,采用四甲基偶氮唑盐微量酶反应比色法(MTT)检测细胞增殖抑制效果,采用流式细胞术检查细胞凋亡情况。结果 MTT结果显示,沙利度胺能够有效的抑制HepG2细胞的增殖,且具有剂量和时间依赖性。作用时间为12h,24h和48h时,沙利度胺浓度为200μg/ml的抑制率最高,与其他各浓度比较,差异有统计学意义(P<0.05)。作用时间为48h时,除50μg/ml外的其他浓度的沙利度胺的抑制率最高,与其他各时间比较,差异有统计学意义(P<0.05)。沙利度胺200μg/ml浓度组和联合组各时间点对HepG2细胞的抑制率比较,差异有统计学意义(P<0.05)。各时间点时,联合组的抑制率高于沙利度胺200μg/ml浓度组和表阿霉素组,差异有统计学意义(P<0.05)。沙利度胺200μg/ml浓度组和联合组的抑制率,48h时最高,与其他各时间点比较,差异有统计学意义(P<0.05)。流式细胞术检验结果显示,作用时间为12h和24h时,沙利度胺梯度组对HepG2细胞凋亡指数(AI)影响的比较,差异有统计学意义(P<0.05)。沙利度胺浓度为50μg/ml、100μg/ml和200μg/ml时,HepG2细胞凋亡指数比较,差异有统计学意义(P<0.05)。各时间点,表阿霉素组、沙利度胺梯度组和联合组对HepG2细胞AI的比较,且联合组的抑制率显著高于其他各组,差异均有统计学意义(均P<0.05)。结论沙利度胺具有体外抑制人肝癌细胞株HepG2增殖和促凋亡作用,具有剂量和时间依赖性,可使用沙利度胺联合表阿霉素治疗肝癌。
Objective To investigate the effects of thalidomide combined with epirubicin on proliferation of human hepatocellular carcinoma cell HepG2 in vitro and to provide basis for the treatment of liver cancer. Methods HepG2 cells were divided into four groups: blank control group, epirubicin group (1.0 mg / L), thalidomide gradient group (50μg / ml, 100μg / ml and 200μg / ml) And 400μg / ml) and combination group (thalidomide 200μg / ml + epirubicin 1.0 mg / L). The effect of 4 groups of drugs on HepG2 cells was observed at 12h, 24h, 48h and 72h. Azadirachtin micro-enzyme reaction colorimetric assay (MTT) inhibition of cell proliferation, flow cytometry was used to detect apoptosis. Results MTT results showed thalidomide could effectively inhibit the proliferation of HepG2 cells in a dose-and time-dependent manner. The inhibitory rate of thalidomide at 200μg / ml was the highest at 12h, 24h and 48h, and the difference was statistically significant (P <0.05) compared with other concentrations. The effect of thalidomide at the concentration of 50μg / ml was the highest at 48h, and the difference was statistically significant compared with other time (P <0.05). Thalidomide 200μg / ml concentration group and the combination group of HepG2 cells at different time points, the difference was statistically significant (P <0.05). At each time point, the inhibition rate of the combination group was higher than that of the thalidomide concentration of 200μg / ml and epirubicin, the difference was statistically significant (P <0.05). The inhibitory rate of thalidomide at the concentration of 200μg / ml and the combination group was the highest at 48h, which was significantly different from other time points (P <0.05). The results of flow cytometry showed that the effect of thalidomide gradient group on apoptosis index (AI) of HepG2 cells was statistically significant (P <0.05) at 12 h and 24 h. The apoptotic index of HepG2 cells was significantly different at thalidomide concentrations of 50μg / ml, 100μg / ml and 200μg / ml (P <0.05). Each time point, epirubicin group, thalidomide gradient group and the combination group on AI of HepG2 cells, and the combined group of inhibition rate was significantly higher than the other groups, the differences were statistically significant (P <0.05 ). Conclusion Thalidomide can inhibit proliferation and induce apoptosis of HepG2 cells in vitro in a dose-and time-dependent manner. Thalidomide and epirubicin can be used to treat HCC.