【摘要】 目的:本实验拟用熊果酸干预处理人急性早幼粒白血病HL-60细胞,体外培养实验,证实其对HL-60细胞的增殖抑制和促进诱导凋亡作用,并通过免疫细胞化学方法检测bcl-2,survivin在熊果酸与该细胞株作用后表达,探讨熊果酸诱导HL-60细胞调亡可能的分子机制。方法:荧光显微镜观察细胞形态;流式细胞仪检测细胞周期相分布及细胞凋亡率;琼脂糖凝胶电泳观察细胞DNA;免疫细胞化学方法检测Bcl-2、Survivin蛋白。结果:荧光显微镜下观察,80μmol/LUA作用24 h的HL-60细胞可看到典型的细胞凋亡形态学变化;40umol/L、80umol/LUA可使G₀/G₁期细胞增多,出现G₀/G₁期阻滞,S期和G₂/M期细胞数减少,凋亡细胞峰（Sub-G₁）及凋亡率逐渐增高,Bcl-2、Survivin蛋白表达降低,当UA浓度为40μmol/L时,P<0.01,有显著性差异。结论:UA呈浓度和时间依赖性抑制HL-60细胞增殖,诱导其凋亡机制可能与下调Bcl-2、Survivin蛋白表达有关。更多还原

【Abstract】 Objective:The experience is to deal with human acute promyelocytic leukemia HL-60 cells with ursolic acid intervention,vitro experiments,confirmed that it had effects on proliferation inhibition and promote induction of apoptosis of HL-60 cells, and detected bcl-2 by immunocytochemistry,survivin express after the UA had an effect on the cell lines,exploring the molecular mechanism which can make ursolic acid to induce apoptosis.Methods:Observed cell morphology by fluorescence microscopy;Detected cell cycle phase distribution and apoptosis rate by flow cytometry;Observed DNA cell with agarose gel electrophoresis;Detected Bcl-2, Survivin protein by immunocytochemistry.Result:Observed morphological changes of apoptosis after 80μmol/L ursolic,acid induced HL-60 cell for 24h;40umol/L,80umol/L UA could increased G₀/G₁ phase cells,appeared G₀/G₁ phase arrest,S phase and G₂/M phase cells decreased,peak of apoptotic cells（Sub-G₁） and the apoptosis rate increased gradually,Bcl-2,survivin protein expression decreased,when the UA concentration for 40μmol/L,P<0.01,there was a significant difference.Conclusion:UA concentration and time inhibition of proliferation of HL-60 cells dependently,apoptosis may be related to decreasing Bcl-2,Survivin protein expression.更多还原