蛋白酶体抑制剂PS-341靶向干预多发性骨髓瘤骨髓间充质干细胞的实验研究

Target Intervention of Proteasome Inhibitor PS-341 on Mesenchymal Stem Cells Derived of Multiple Myeloma

目的:分析蛋白酶体抑制剂PS-341对多发性骨髓瘤(MM)骨髓间充质干细胞(MSCs)凋亡及其对IL-6、IL-1β、SCF细胞因子表达的影响。方法:含10%FBS的1640培养液培养得到MSCs,流式细胞术(FCM)鉴定所培养细胞的表型,以终浓度分别为0 nmol/L、50 nmol/L、100 nmol/L、150 nmol/L、200 nmol/L的PS-341作用MSCs 4 h,光学显微镜观察细胞形态改变,荧光显微镜下Annexin V-FITC/PI双染法观察细胞凋亡,反转录聚合酶链反应(RT-PCR)检测IL-6、IL-1β、SCF细胞因子表达的变化。结果:PS-341可诱导MSCs凋亡,与其浓度成正比;与对照组(0 nmol/L PS-341)比较,浓度分别为50 nmol/L、100 nmol/L、150 nmol/L、200 nmol/L的PS-341作用MSCs 4 h后明显抑制IL-6、IL-1β、SCF的表达(P<0.05);PS-341对IL-6和SCF的抑制与PS-341浓度成正相关(P<0.05),在初发/难治复发和完全缓解(CR)两组患者间均无显著差异;对IL-1β的抑制在CR组较初发/难治复发组更明显(P<0.05);PS-341作用后IL-1β表达的强弱对IL-6及SCF的表达无影响。结论:PS-341诱导MM患者MSCs凋亡和抑制其IL-6、IL-1β、SCF的表达,是PS-341有效治疗MM的靶点之一。

Objective.To analyse the apoptosis and expressions of cytokines IL-6,IL-1β and SCF of mesenchymal stem cells（MSCs）,which was derived from Multiple Myeloma （MM） treated with proteasome inhibitor PS-341. Meth ods ：MSCs were incubated in the culture of RPMI1640 with 10% fetal bovine serum（FBS） and were determined by flow cytometric （FCM）,then MSCs were exposed to 0 nmol/L ,50 nmol/L, 100 nmol/L, 150 nmol/L ,200 nmol/L PS-341 for 4 hours,then cells apoptosis were analyzed by inverted light microscopy and fluorescence microscope with annexin VFITC/PI dual staining,and reverse transcription polymerse chain reaction （RT-PCR） was performed to detect the expressions of IL-6,IL-1β and SCF. Results： PS-341 induced MSCs apoptosis in a dose-dependent manner. After PS-341 treated expressions of IL-6,IL-1β,SCF of MSCs were decreased（P〈0.05）,There were no statistics differences of IL-6 and SCF expression between refractory/relapsed group and complete remission（CR）group and PS-341 downregulated the expressions of IL-6 and SCF of MSCs in a dose-dependent manner;IL-1β was significantly inhibited in CR group compared with that in refractory/relapsed group; there were no significant relationship between the expressions of IL-6 and SCF and the expression of IL-1β. Conclusions ：Proteasome inhibitor PS-341 can induce MSCs apoptosis and inhibited the expressions of IL-6,IL-1β and SCF,which might be one of mechanisms and targets for MM treatment.