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本研究探讨2-甲氧基雌二醇(2-ME)对慢性髓系白血病(CML)K562细胞caspase-3和survivin表达的影响。实验分3组:对照组,培养基中不含2-ME;实验组,分别用1、2、4、8及16μmol/L的2-ME处理K562细胞;阴性对照组,培养液中以不含RNase的灭菌蒸馏水代替K562细胞。应用TUNEL、流式细胞术(FCM)、半定量RT-PCR分别检测K562细胞凋亡率、caspase-3及survivin蛋白及其基因表达水平。结果表明:2-ME在一定的浓度范围内呈剂量依赖形式诱导K562细胞凋亡,与对照组比较差异均有统计学意义(p均<0.01),且TUNEL和FCM方法检测的K562细胞凋亡率呈正相关(γ=0.845,p=0.034)。随着2-ME浓度增加,caspase-3蛋白表达量逐渐增加,而survivin蛋白表达量逐渐降低,两者与对照组比较差异均有统计学意义(p均<0.05);且caspase-3与survivin蛋白表达量之间呈负相关(γ=-0.956,p=0.001)。随着2-ME浓度增加,caspase-3基因表达量逐渐增加,而survivin基因表达量逐渐降低,两者与对照组比较差异均有统计学意义(p均<0.01),且caspase-3与survivin基因表达量之间呈负相关(γ=-0.966,p=0.001)。结论:2-ME能够以剂量依赖形式诱导K562细胞凋亡,显示了其对CML有潜在的治疗价值。
This study was aimed to investigate the effect of 2-methoxyestradiol (2-ME) on the expression of caspase-3 and survivin in chronic myeloid leukemia (CML) K562 cells. The experiment was divided into three groups: the control group, the medium without 2-ME; the experimental group, respectively, with 1, 2, 4, 8 and 16μmol / L 2-ME treatment of K562 cells; negative control group, Sterile distilled water containing RNase instead of K562 cells. TUNEL, FCM and semi-quantitative RT-PCR were used to detect the apoptosis rate, the expression of caspase-3 and survivin protein and its gene expression in K562 cells. The results showed that 2-ME induced apoptosis in K562 cells in a dose-dependent manner in a dose-dependent manner, with significant difference compared with the control group (all p <0.01), and apoptosis of K562 cells detected by TUNEL and FCM The rate was positively correlated (γ = 0.845, p = 0.034). With the increase of 2-ME concentration, the expression of caspase-3 protein gradually increased, while the expression of survivin protein gradually decreased. There was significant difference between the two groups (P <0.05), and the expression of caspase-3 and survivin Protein expression was negatively correlated (γ = -0.956, p = 0.001). With the increase of 2-ME concentration, the expression of caspase-3 gene gradually increased while the expression of survivin gene gradually decreased. There was significant difference between the two groups (all p <0.01), and the expressions of caspase-3 and survivin Gene expression was negatively correlated (γ = -0.966, p = 0.001). Conclusion: 2-ME can induce apoptosis of K562 cells in a dose-dependent manner, which shows its potential therapeutic value for CML.