摘要
目的:研究持续低剂量磷酰胺氮芥(phosphoramide,PM)联合泼尼松龙(prednisolone)(即CP节律性化疗方案)对骨髓瘤细胞RPMI 8226增殖及凋亡的影响,并探讨其对Notch1/NF-κB信号通路的调控作用。方法:实验分DMSO对照组,磷酰胺氮芥(PM)组,泼尼松龙组,磷酰胺氮芥+泼尼松龙组(即CP组),采用不同组的药物处理骨髓瘤细胞RPMI 8226,用CCK8法检测细胞增殖,流式细胞术检测细胞周期及凋亡,RT-PCR检测Notch1和NF-κB的mRNA表达水平。结果:与DMSO对照组相比,随着时间的延长,PM、泼尼松龙及CP组均使RPMI 8226细胞的增殖抑制率明显提高(r分别为0.994、0.996、0.999,P<0.001),并且在相同作用时间,不同组的药物对细胞的增殖抑制率也明显不同,PM对细胞的抑制率最弱,而CP组对细胞的抑制率最强,泼尼松龙组次之(P<0.01)。用药处理48 h后,与DMSO对照组相比,各用药组G_1/G_0期细胞比例明显增加,S期细胞比例明显减少,尤其在PM和CP组,G2/M期细胞比例在PM增加,而在泼尼松龙和CP组则减少。用药处理48 h后,与DM SO对照组相比,治疗组PM、泼尼松龙、CP组的RPMI 8226细胞凋亡率依次增高(P<0.01)。用药处理48 h后,与DMSO对照组相比,泼尼松龙组、PM组、CP组RPMI 8226细胞的Notch1和NF-κB mRNA表达均依次明显下降(P<0.001)。结论:CP节律性化疗可以显著减少RPMI 8226细胞增殖,促进其凋亡,使RPMI 8226细胞主要阻滞于G_1/G_0期,并能显著降低Notch1和NF-κB的表达水平,显示CP节律性化疗对MM的治疗作用有Notch1/NF-κB信号通路的参与。
Objective：To investigate the effect of metronomic chemotherapy of low dose phosphoramide combined with prednisolone（CP metronomic chemotherapy） on proliferation and apoptosis of RPMI 8226 cells,and to explore its regulating effect on Notch1/NF-κB signaling pathways.Methods：Experiment was divided into the DMSO control group,and the phosphoramide mustard（PM） group,the prednisolone group,the phosphoramide mustard plus prednisolone group（the CP group）.RPMI 8226 cells were treated with different drugs,CCK-8 method was used to detect cell proliferation,flow cytometry was used to detect the cell cycle and apoptosis,reverse transcription PCR was used to detect Notchl and NF-κB mRNA expression level.Results：Compared with DMSO control group,RPMI8226 cell proliferation inhibition rate in all the PM,prednisolone and CP groups increased significantly with prolonging of time（r of 0.994,0.996,0.999,respectively,P〈0.001）.And at the same time,the inhibitory rate of cell proliferation was significantly different;the cell inhibitory rate in PM group was lowest,that in CP group was highgest,that in prednissone group was intermediate（P〈0.01）.After 48 hours,compared with the DMSO control group,the G1/G0cell proportion in treatment group increased significantly,S phase cell proportion decreased significantly,especially in PM and CP groups.The G2/M phase cell proportion increased in PM group,while reduced in the prednisolone and the CP groups.After 48 hours,compared with the DMSO control group,RPMI 8226 cell apoptosis rate increased as follow：in PM,pre-dnisolone and CP group（P〈0.01）.After 48 hours,compared with the DMSO control group,Notchl and NF-κB mRNA expression in the prednisolone,the PM and the CP group decreased significantly（P〈0.001）.Conclusion：CP metronomic chemotherapy can significantly reduce RPMI 8226 cell proliferation,promote RPMI 8226 cell apoptosis,arrest RPMI 8226 cells mainly in the G1/G0 phase,and significantly reduce Notchl and NF-κB expression levels.It is suggested that Notchl /NF-κB signaling pathways is involved in CP metronomic chemotherapy for MM.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2016年第5期1470-1475,共6页
Journal of Experimental Hematology
基金
上海市卫生局计划项目(20134411)
