Embelin通过上调死亡受体DR4/DR5mRNA表达增加HL-60细胞对TRAIL的敏感性

Embelin sensitizes HL- 60 cells to TRAIL by upregulation of DR4 /DR5 mRNA expression

目的:探讨Embelin增加HL-60细胞对肿瘤坏死因子相关凋亡诱导配体(TRAIL)的敏感性及可能的作用机制。方法:TRAIL 1、5、10、50及100 ng/ml分别处理HL-60细胞6、12、24及48h,MTT法绘制细胞生长曲线;TRAIL 10 ng/ml±亚细胞毒浓度的Embelin处理HL-60细胞6、12、24及48h,Annexin V/PI复染流式细胞术检测细胞凋亡,Western blot检测24h时Caspase-3、8及9的表达;亚细胞毒浓度的Embelin处理HL-60细胞6、12、24及48h,实时荧光定量PCR检测DR4及DR5 mRNA的表达。结果:TRAIL对HL-60细胞具有增殖抑制作用。亚细胞毒浓度的Embelin联合10 ng/ml TRAIL作用于HL-60细胞时细胞凋亡率较单独应用TRAIL时增加,相应的Caspase-3、8及9的表达也随之增加。结论:亚细胞毒浓度的Embelin可以增加HL-60细胞对TRAIL的敏感性,其机制可能与上调DR4及DR5 mRNA表达并进而启动凋亡途径有关。

Objective：To investigate whether Embelin could sensitize HL-60 cells to TRAIL or not and explore the possible mechanism. Methods：HL- 60 cells were treated with TRAIL of 1,5,10,50 and 100 ng/ml for 6,12,24 and 48h seperatedly and proliferation curve was plotted by MTT assay. HL - 60 cells were treated with 10ng/ml TRAIL with or without Embelin of subtoxic concentration for 6,12,24 and 48h seperatedly,and cell apoptosis rate was detected by Annexin V/PI double staining and flow cytometry and Caspase - 3,8 and 9 at 24h by Western blot. HL - 60 cells were treated with Embelin of subtoxic concentration for 6,12,24 and 48h seperatedly,and DR4 and DR5 mR- NA were detected by real - time PCR. Results：TRAiL inhibited HL - 60 cells proliferation. TRAIL combined with Embelin of subtoxic concentration was more effective in inducing cell apoptosis than TRAIL alone and Caspase - 3,8 and 9 expression increase correspondingly. Conclusion： Embelin of subtoxic concentration could sensitize HL -60 cells to TRAIL. Upregulation of DR4 and DR5 mRNA and initiation of apoptosis pathway may contribute to the possi- ble mechanism.