蛇床子素对TRAIL诱导白血病HL-60细胞凋亡的作用及其相关机制研究

Effects of Osthole on Apoptosis of HL-60 Cells Induced by TRAIL and Its Related Mechanism

目的:探讨蛇床子素(Osthole)对肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导白血病HL-60细胞凋亡的影响及其可能机制。方法:采用MTT法检测不同浓度Osthole和TRAIL单独及联合应用对HL-60细胞增殖的影响。选择低于半数抑制浓度(IC50)的100μmol/L的Osthole和40 ng/ml TRAIL单独或联合处理细胞,通过流式细胞术测定HL-60细胞凋亡和细胞线粒体膜电位(MMP)的变化,RT-PCR法检测BCL-2、BAX和DR5 mRNA的表达,用分光光度法检测Caspase-3、-8、-9活性变化。结果:100μmol/L Osthole和40 ng/ml TRAIL联合处理HL-60细胞48 h,HL-60细胞的凋亡率达(33.9±2.7)%,较单用Osthole、TRAIL均显著提高(P<0.05),同时2者联合应用能增强降低MMP和BCL-2/BAX表达比值的效果,提高DR5的表达和Caspase-3、-8、-9活性。结论:Osthole能增强TRAIL诱导HL-60细胞凋亡的敏感性,其机制可能与其激活线粒体凋亡途径和上调DR5的表达密切相关。

Objective： To explore the effects of Osthole on apoptosis of HL-60 cells induced by tumor necrosis factor related apoptosis inducing ligand（ TRAIL） and its possible mechanism. Methods： The proliferative inhibition of HL-60 cells treated with different concentrations of Osthole,TRAIL alone and Osthole combined with TRAIL was measured by MTT assay. The HL-60 cells were treated with Osthole,TRAIL alone and Osthole combined with TRAIL at the concentration IC50 value,i. e. 100 μmol/L for Osthole and 40 ng/ml for TRAIL. Apoptosis and mitochondrial membrane potential（ MMP） of HL-60 cells were detected by flow cytometry; the mRNA expression of BCL-2,BAX and DR5 was determined by RT-PCR; and the levels of Caspase-3,-8,-9 activity were detected by spectrophotometry.Results： The combined treatment（ 100 μmol/L Osthole ＋ 40 ng/ml TRAIL） of HL-60 cells for 48 h induced an apoptotic rate of（ 33. 9 ± 2. 7） %,which was significantly higher than that of cells treated with Osthole or TRAIL alone（ P〈0. 05）; at the same time,the combined treatment promoted the decrease of MMP and the expression rate of BCL-2/BAX,and potentiated the expression of DR5 and Caspase-3,-8,-9 activity. Conclusion： Osthole can sensitize HL-60 cells to TRAIL-induced apoptosis,which may be related with the activation of mitochondrial pathways and up-regulation of DR5.