丙戊酸钠对多发性骨髓瘤RPMI 8226细胞Notch通路的影响  

Effects of VPA on the Expression of Notch Signaling Pathway in Multiple Myeloma RPMI 8226 Cell Line

作　　者：乔莲[1] 杨悦[2] 赵瑞娟[2] 王丽红[2] 赵蕾[2] 闫丽娜[2] 张志华[2] 郝长来[2] 

机构地区：[1]河北中医学院,河北石家庄050200 [2]承德医学院附属医院血液科,河北承德067000

出　　处：《中国实验血液学杂志》2016年第5期1449-1453,共5页Journal of Experimental Hematology

基　　金：河北省自然科学基金项目(H2013406112)

摘　　要：目的:探讨丙戊酸钠(VPA)对多发性骨髓瘤RPMI 8226细胞Notch通路中Notch1受体活性片段ICN1和靶基因Hes1表达的影响。方法:实验分为4组:空白对照组、VPA 2、4和8 mmol/L组,应用四甲基偶氮唑蓝(MTT)检测VPA对多发性骨髓瘤RPMI 8226细胞的增殖抑制作用,采用反转录聚合酶链式反应(RT-PCR)检测Notch通路中Notch1受体活性片段ICN1、靶基因Hes1 mRNA的表达,应用蛋白印记法(Western blot)检测ICN1、Hes1蛋白的表达。结果:同一时间、不同浓度VPA(0、2、4、8 mmol/L)处理RPM I 8226细胞,其细胞的生长明显受到抑制。相同浓度的VPA作用于RPMI 8226细胞不同时间(24、48、72 h)时,细胞的生长明显受到抑制。2、4、8mmol/L VPA处理48 h时与空白对照组相比,ICN1 mRNA及蛋白表达水平均明显降低(P<0.05),靶基因Hes1mRNA及蛋白表达水平均明显降低,(P<0.05)。结论:VPA明显抑制多发性骨髓瘤RPM I 8226细胞的增殖,并且这种抑制作用在一定范围内呈时间-浓度依赖性(r=0.945)。VPA可以下调Notch通路中Notch1受体活性片段ICN1、靶基因Hes1 mRNA及蛋白的表达水平,VPA可能通过抑制Notch信号通路实现对多发性骨髓瘤细胞的生长抑制作用。Objective：To investigate the effects of valproic acid（VPA） on the expression of intracellular domain of Notchl（ICN1） and Hesl in multiple myeloma RPMI 8226 cell line.Methods：Experiments were divided into 4 group：blank control group and groups of cells treated with VPA of different concentration（2,4,8 mmol/L）,the cell proliferation was detected by MTT method,RT-PCR was applied to detect the mRNA expression level of ICN1 and Hesl.Western blot was used to detect the protein expression of ICN1 and Hesl.Results：The proliferation of the RPMI8226 cell was obviously inhibited by different concentration of VPA（2,4,8 mmol/L） at the same time.The same concentration of VPA was used to treat RPM18226 cell for different time（24,48,72 h）,the cell proliferation was obviously inhibited.Compared with control group,the mRNA and protein expression of ICN1 was significantly depressed at different concentration of VPA（2,4,8 mmol/L） for 48 h（P〈0.05）.Compared with control group,the mRNA and protein expression of Hes1 was depressed at different concentration 2,4,8 mmol/L） of VPA for 48 h（P〈0.05）.Conclusion：VPA inhibits the proliferation of the RPMI 8226 cell in a time-and dose-dependent manner;VPA down-regulates the mRNA and protein expression level of ICN1 and Hesl in RPMI8226 cell;thus VPA might inhibit cell proliferation possibly through the inhibition of Notch signaling pathway in multiple myeloma cells.

关 键 词：多发性骨髓瘤 丙戊酸钠 NOTCH信号通路 ICN1 HES1 

分 类 号：R733.3[医药卫生—肿瘤]