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目的观察表柔比星(EPI)温热化疗对人胃癌MGC803细胞的体外抑制及凋亡的影响,探讨其作用机制。方法采用四氮甲唑蓝(MTT)法确定作用48 h抑制率为50%的药物浓度作为实验的工作浓度。以48 h的IC50的药物浓度进行化疗或与热疗的联合,热疗选择温度为43℃,体外作用于MGC803细胞,实验分四组:对照组(C)、热疗组(H)、化疗组(D)、热化疗组(HD)。MTT法检测各处理组对肿瘤细胞增殖的抑制作用;应用RT-PCR观察各处理组诱导MGC803细胞凋亡的作用及对Caspase-3和Bcl-2表达变化的影响。结果单纯热疗和化疗对MGC803细胞系均有抑制作用(P<0.05);EPI温热化疗可明显增强EPI对MGC803细胞的抑制作用(P<0.05);与对照组比,热疗组、化疗组及热化疗组细胞凋亡率均较对照组显著升高,各组之间均有差异(P<0.05);Bcl-2表达下调、Caspase-3表达上调,各组之间均有差异(P<0.05)。Caspase-3与Bcl-2蛋白的表达呈明显负相关(r=-0.990,n=12),细胞凋亡率与Bcl-2蛋白表达负相关(r=-0.924,n=12),而与Caspase-3蛋白表达正相关(r=0.891,n=12)。结论 EPI联合热疗能显著抑制MGC803细胞生长并诱导细胞凋亡,可能与下调Bcl-2,上调Caspase-3的表达及二者相互作用有关。
Objective To observe the effects of epirubicin (EPI) chemotherapy on human gastric cancer cell line MGC803 in vitro and its mechanism of action. Methods MTT method was used to determine the concentration of the drug whose inhibitory rate was 50% after 48 h treatment as experimental concentration. Chemotherapy or hyperthermia after 48 h at IC50 concentrations of the drug combination, hyperthermia select the temperature of 43 ℃, MGC803 cells in vitro, were divided into four groups: control group (C), hyperthermia group (H), chemotherapy Group (D), thermochemotherapy group (HD). Inhibition of each treatment on tumor cell proliferation by MTT assay; by RT-PCR was observed in each treatment group MGC803 cells inducing apoptosis of change and its effect on Caspase-3 and Bcl-2 expression. Results hyperthermia and chemotherapy alone MGC803 cell lines were inhibited (P <0.05); EPI was warmed chemotherapy can significantly enhance the inhibitory effect of EPI MGC803 cells (P <0.05); and the control group, group hyperthermia, chemotherapy (P <0.05). The expressions of Bcl-2 and Caspase-3 were up-regulated in all groups (P <0.05) P <0.05). There was a negative correlation between the expression of Caspase-3 and Bcl-2 protein (r = -0.990, n = 12), and the apoptosis rate was negatively correlated with the expression of Bcl- Caspase-3 protein expression was positively correlated (r = 0.891, n = 12). Conclusion EPI combined with hyperthermia can significantly inhibit MGC803 cell growth and induce apoptosis, which may be related to the down-regulation of Bcl-2, up-regulation of Caspase-3 and their interaction.