骨髓瘤U266细胞中Blimp1与内质网应激ATF4/CHOP信号通路的关联性研究  

Correlation of Blimp1 with ATF4/CHOP Signaling Pathway in Multiple Myeloma U266 Cells

作　　者：刘俊文 徐锦 刘红春[2] LIU Jun-Wen;XU Jin;LIU Hong-Chun(Department of Clinical Laboratorial Examination.The Children′s Hospital of Fudan University,Shanghai 201102,China;Department of Clinical Laboratorial Examination,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan Province,China)

机构地区：[1]复旦大学附属儿科医院临床检验中心,上海201102 [2]郑州大学第一附属医院检验科,河南郑州450052

出　　处：《中国实验血液学杂志》2020年第4期1215-1220,共6页Journal of Experimental Hematology

基　　金：河南省教育厅科技研究重点项目(14A320026);河南省卫生厅医学科技攻关重点项目(201402008);河南省科技厅攻关项目(172102310028)。

摘　　要：目的:探讨Blimp1是否通过干扰内质网应激诱导的ATF4/CHOP细胞凋亡通路在骨髓瘤中发挥抗凋亡作用,以及探讨阿司匹林是否通过抑制Blimp1表达发挥抗肿瘤作用。方法:收集初诊骨髓瘤患者(40例)和增生骨髓象者(30例)的骨髓液,流式细胞术分选表型异常及正常的浆细胞,LV-Blimp1-RNAi(40051-2)重组慢病毒下调U266细胞株的Blimp1表达,并检测ATF4和CHOP基因表达的变化。不同浓度的阿司匹林溶液体外刺激U266细胞株,CCK-8检测细胞的增殖活性,real-time PCR检测Blimp1,ATF4和CHOP表达水平。结果:表型异常的浆细胞Blimp1表达水平较正常浆细胞显著升高,ATF4和CHOP表达水平显著降低,差异均有统计学意义。在MOI=100的条件下,慢病毒表达载体感染U266细胞,荧光显微镜下观察显示转染效率>80%。阳性实验组与空白对照组和阴性对照组比较,Blimp1表达水平显著下调,ATF4和CHOP表达水平显著升高,差异均有统计学意义。CCK-8结果显示,阿司匹林可显著抑制U266骨髓瘤细胞的增殖活性,抑制作用随药物刺激时间的延长和刺激浓度的增加而增强,同时,U266细胞中Blimp1的表达水平随阿司匹林浓度升高而降低,而ATF4和CHOP的表达水平随药物浓度升高而升高。结论:U266骨髓瘤细胞中Blimp1可能通过干扰ATF4/CHOP信号通路发挥抗凋亡作用。小剂量阿司匹林可能通过抑制骨髓瘤细胞中Blimp1表达发挥抗肿瘤活性。Objective:To investigate whether Blimp1 plays an anti-apoptosis role in myeloma by interfering with ATF4/CHOP cell apoptosis pathway induced by endoplasmic reticulum stress,and to explore the anti-myeloma mechanism of aspirin.Methods:The bone marrow fluid of 40 newly diagnosed multiple myeloma patients without treatment and 30 control people with relatively normal bone marrow was collected.Flow cytometry was used to separated the normal and abnormal plasma cells,LV-Blimp1-RNAi(40051-2)recombinant lentivirus down-regulates the expression of Blimp-1 in U266 cell line and detected the changes of the expression of ATF4 and CHOP gene.U266 cells were stimulated by aspirin at different concentrations(0,0.5,2.5,5.0 mmol/L)in vitro.Then the effect of aspirin on proliferation of U266 cells was measured by CCK-8 assay,the mRNA expression levels of Blimp1,ATF4 and CHOP in four groups were detected by real-time PCR.Results:The expression level of Blimp1 in phenotype abnormal plasma cells was significantly increased as compared with normal cells,while the expression of ATF4 and CHOP in phenotype abnormal plasma cells was significantly decreased as compared with normal cells(P<0.05).In the case of MOI=100,the transfection efficiency of U266 cells was beyond 80%as detected by fluorescence microscopy.Compared with blank conrol and negatine control groups,Blimp1 mRNA expression level in positive group was significantly reduced while ATF4 and CHOP expression significantly increased.CCK-8 showed that the proliferation activity of U266 cells could be inhibited by aspirin,which showed a time-and dose-dependent manner;at the same time,the expression level of Blimp1 in U266 cells were decreased with the increasing of aspirin concentration,while the expression level of ATF4 and CHOP was increased with the increasing of aspirin concentration.Conclusions:Blimp1 may display the anti-apoptosis of myeloma cells through interfering with ATF4/CHOP signaling pathway;low dose of aspirin may play anti-myeloma effect by inhibiting the expression of

关 键 词：多发性骨髓瘤 Blimp1 ATF4 CHOP 阿司匹林 

分 类 号：R733.3[医药卫生—肿瘤]