蛋白酶体抑制剂PS-341对U266细胞凋亡和多种细胞因子表达影响的研究

Study on the effects of proteasome inhibitor PS-341 on apoptosis and expression of cytokines of U266 cells

目的：探讨蛋白酶体抑制剂PS-341（bortezomib）诱导多发性骨髓瘤细胞株U266细胞凋亡时对IL-6、IL-1β和SCF细胞因子表达的影响。方法：用含15％FBS的RPM11640培养液体外培养U266细胞，分别用0、50、100、150、200nmol／L的PS-341干预U266细胞4h后收集，荧光显微镜观察细胞凋亡，Annexin V-FITC／PI双染法流式细胞术（FCM）检测细胞凋亡率的变化，反转录聚合酶链反应（RT-PCR）分析U266细胞IL-6、IL-1β和SCF三种细胞因子表达的改变。结果：荧光显微镜和FCM发现PS-341以浓度依赖方式诱导U266细胞凋亡；PS-341明显抑制U266细胞IL-6、IL-1β和SCF的表达（P〈0．05）；对IL-6和SCF的抑制与PS-341浓度成正相关，对IL-1β的抑制在PS341浓度为0～150nmol／L时成正比关系，当浓度大于150nmol／L时差异无显著性。结论：蛋白酶体抑制剂PS-341诱导U266细胞凋亡时明显抑制其IL6、IL-1β和SCF的表达，是有效治疗MM的机制之一。

Objective：To investigate the effects of proteasome inhibitor PS-341 （Bortezomib） on the expression of interleukin-6 （IL-6） ,interleukin-1β（IL-1β） and stem cell faetor（SCF） and the apoptosis of U266 cells. Methods：U266 Cells were incubated with RPMI1640 and exposed to 0,50,100,150,200 nmol/L of PS-341 for 4 h, respectively. Then the apoptosis of ceils was analyzed by fluorescence microscope and flow cytometry （FCM） with Annexin V-FITC/PI double staining, and the expression of IL-6, IL-1β and SCF was detected by RT-PCR. Results： Fluorescence microscope and FCM showed that PS-341 induced U266 ceils apoptosis in a dose-dependent manner;, The expression of IL-6, IL-1β and SCF was decreased significantly after PS-341 treatment. The inhibitory effect on IL-6 and SCF was positively related with drug concentration. The expression of IL-1β was decreased gradually when the drug concentration was increased from 0 nmol/L to 150 nmol/L, whereas the expression level of IL-1β remained unchangeable when the drug concentration exceeded 150 nmol/L Conclusion：Proteasome inhibitor PS-341 could significantly downregulate the expressions of IL-6 ,IL-1β and SCF in U266 cells, which might be one of the mechanisms for PS-341 for treatment of multiple myeloma.