eIF4E对多发性骨髓瘤CD138^+细胞自噬的影响

Effect of eIF4E on Autophagy of CD138^+ Cells in Multiple Myeloma

目的:探讨真核细胞翻译起始因子4E(eIF4E)对多发性骨髓瘤CD138^+浆细胞自噬的影响。方法:eIF4E抑制剂4EGI处理多发性骨髓瘤CD138^+浆细胞,采用荧光定量PCR和Western blot法检测细胞自噬相关因子LC3-Ⅱ和Beclin1的变化,MTT法检测细胞增殖抑制的变化,流式细胞术检测细胞凋亡。结果:荧光定量PCR检测显示,4EGI处理骨髓瘤细胞后,LC3-Ⅱ、Beclin1 mRNA的表达水平具有时间及浓度依赖性,随着4EGI浓度增加和作用时间延长,LC3-Ⅱ、Beclin1 mRNA的表达水平逐渐增加,差异均有统计学意义(48 h:LC3-Ⅱ,r=0.942,Beclin1,r=0.952;80μg/ml:LC3-Ⅱ,r=0.966,Beclin1,r=0.998)。Western blot检测显示,随着4EGI浓度增加,LC3-Ⅱ、Beclin1蛋白表达逐渐增加(LC3-Ⅱ,r=0.923;Beclin1,r=0.977)。CCK-8检测显示,细胞抑制率逐渐增加(r=0.996);4EGI处理各组(20、40、80μg/ml组)细胞凋亡率(23.23±4.47、7.59±1.67、2.03±0.19)均明显高于对照组(0.03±0.04)(P<0.05)。结论:抑制eIF4E对多发性骨髓瘤CD138^+浆细胞的自噬有激活作用,并引起骨髓瘤细胞自噬性死亡。

Objective:To investigate the effect of eukaryotic translation initiation factor 4 E(eIF4E)on the autophagy of CD138^+plasma cells in multiple myeloma(MM).Methods:Multiple myeloma CD138^+plasma cells were treated with eIF4 E inhibitor 4 EGI,the changes of autophagy-related factors LC3-Ⅱand Beclin1 were detected by fluorescent quantitative PCR and Western blot,the changes of cell proliferation inhibition were detected by MTT assay,and cell apoptosis was detected by flow cytometry.Results:Quantitative fluorescence PCR showed that after treatment of myeloma cells with 4 EGI,the expression levels of LC3-Ⅱand Beclin1 mRNA gradually increased with the enhancomer of 4 EGI concentration and the prolongation of action time,and the differences were statistically significant(48 h:LC3-Ⅱ,r=0.942,Beclin1,r=0.952;80μg/ml:LC3-Ⅱ,r=0.966,Beclin1,r=0.998);Western blot showed that with the enhancement of 4 EGI concentration,the expression of LC3-Ⅱand Beclin1 protein gradually increased(LC3-Ⅱ,r=0.923,Beclin1,r=0.977);CCK-8 showed that the inhibition rate of cells gradually increased(r=0.996);the apoptotic rate of 4 EGI-treated groups(23.23±4.47,7.59±1.67,2.03±0.19)was significantly different from that of control group(0.03±0.04)(P<0.05).Conclusion:The inhibition of eIF4 E can activate the autophagy of CD138^+plasma cells in multiple myeloma and induce the death of myeloma cells.