冬凌草甲素对HL-60细胞凋亡及Survivin表达的影响

Effects of rebescensine A on HL-60 cell apoptosis and expression of Survivin

目的探讨冬凌草甲素对人白血病细胞株HL-60细胞生长抑制作用及其机制。方法收集经不同浓度冬凌草甲素干预的HL-60细胞，制成细胞涂片，光学显微镜下观察细胞形态。采用MTT比色法观察不同浓度冬凌草甲素对HL-60细胞增殖的抑制作用，通过凋亡试剂盒AnnexinV—FITC标记，流式细胞仪检测细胞凋亡率，并以RT-PCR检测凋亡抑制基因Survivin的表达。结果冬凌草甲素在10-25μmol／L范围内可抑制HL-60细胞的生长，且呈时间和浓度依赖性。并可诱导HL-60细胞发生凋亡，表现出特异性的凋亡形态学改变。流式细胞仪分析显示，冬凌草甲素处理后凋亡细胞比例增多，且随着药物浓度的增加而逐渐增加。RT-PCR检测结果显示，20μmol／L冬凌草甲素作用后，与对照组相比，HL-60细胞Survivin mRNA水平下降，且随时间延长表现得更为明显。结论冬凌草甲素能抑制人白血病细胞株HL-60细胞的生长增殖，并可诱导其发生凋亡，其作用机制可能与凋亡抑制基因Survivin的下调有关。

Objective To investigate the anti-proliferation effect of rebescensine A on leukemia HL-60 cells and its mechanism. Methods HL-60 cells in culture medium in vitro were given different concentrations of rebescensine A. Morphological changes were observed in cell smears. MTT assay was used to observe the growth inhibition of HL-60 cells ; Annexin V-FITC and flow cytometry were used to examine cell apoptosis rate. RT-PCR was used to observe the expression of Survivin. Results Rebescensine A （ 10-25 μmol/ L} could inhibit the growth of HL-60 cells and induce apoptosis significantly in a time- and dose-dependent manner. Marked morphological changes of cell apoptosis were observed. Apoptotic cell count increased along with increasing concentrations of rebescensine A. The down-regulated expression of Survivin mRNA was showed by RT-PCR and changed more obviously with prolongation in HL-60 cells treated by rebescensine A 20 μmoL/L. Conclusion Rebescensine A could inhibit proliferation and induce apoptosis of HL-60 cells in vitro. One of the mechanisms may be the down-regulated expression of survivin mRNA.