熊果酸诱导HL-60细胞凋亡机制的探讨

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，80gmol／LUA作用24h的HL一60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40gmol／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。

An exploration on mechanism inducing apoptosis of HL-60 cells with ursolic acid

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 ceils, 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 80ttmol/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 40pmol/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.