靶向抑制miR-21对多发性骨髓瘤细胞凋亡、增殖的影响及其作用机制的初步研究  

The influence of reduced miR-21 expression on proliferation and apoptosis in multiple myeloma cell and its mechanism

作　　者：宋慧慧[1,2] 李原 马钰 凌孙凯歌[1] 葛峥 黄培林 SONG Huihui;LI Yuan;MA Yu;LING Sunkaige;GE Zheng;HUANG Peilin(School of Medicine,Southeast University,Nanjing 210009,China;Department of Hematology,Zhongda Hospital, Southeast University,Nanjing 210009,China)

机构地区：[1]东南大学医学院,江苏南京210009 [2]东南大学附属中大医院血液内科,江苏南京210009

出　　处：《东南大学学报（医学版）》2018年第5期819-823,共5页Journal of Southeast University(Medical Science Edition)

摘　　要：目的:研究miR-21对多发性骨髓瘤细胞增殖及凋亡产生的作用,初步探讨其作用机制。方法:qRTPCR法检测miR-21在多发性骨髓瘤各细胞株中的表达水平。将miR-21抑制物及其阴性对照物转染至U266细胞株,RT-PCR法检测U266细胞miR-21的表达变化,CCK-8法检测miR-21对U266细胞增殖的影响,流式细胞术分析miR-21对U266细胞凋亡的作用,Western blotting检测U266细胞中PETN及FOXO1蛋白表达水平。结果:转染后实验组miR-21相对表达水平为0. 286±0. 031,低于阴性对照组(1. 015±0. 14)(P <0. 05)。实验组U266细胞的增殖抑制率为(53. 04±0. 02)%,显著高于阴性对照组[(20. 89±0. 17)%](P <0. 05);流式分析显示实验组U266细胞比阴性对照组凋亡明显增加[(28. 53±3. 23)%vs(5. 12±0. 63)%],差异具有统计学意义(P <0. 05)。Western blotting结果显示,与阴性对照组、空白对照组相比,实验组U266细胞的PTEN、FOXO1蛋白表达水平均上调(P <0. 05)。结论:靶向抑制miR-21可以促进多发性骨髓瘤细胞凋亡,抑制细胞增殖,这种作用可能是通过上调PTEN和FOXO1水平来实现的。Objective:To explore the effect of miR-21 on proliferation and apoptosis of multiple myeloma cell lines and its molecular mechanism.Methods:miR-21 was detected on multiple myeloma(MM)cell lines by qRT-PCR.U266 cell line was transfected with miR-21 inhibitor and its negative control.qRT-PCR was used to detect the miR-21 expression changes.Cell proliferation and apoptosis were determined by using CCK-8 and flow cytometry.Western blotting was used to detect the protein expression of PTEN and FOXO1.Results:The relative expression of miR-21 in experimental group was 0.286±0.031 after transfection,which was lower than that in negative control group(P<0.05).The proliferation inhibition rate was(53.04±0.02)%in experimental group,which was down to(20.89±0.17)%in negative control group(P<0.05).The cellular apoptotic rate in the experimental group was significantly higher than that in the negative control group[(28.53±3.23)%vs(5.12±0.63)%](P<0.05).Western blotting analysis revealed up-regulation of PTEN and FOXO1 protein expression in experimental group by inhibiting miR-21 expression(P<0.05).Conclusion:Inhibiting miR-21 expression has distinct effects on apoptosis and proliferation inhibition,which is mediated by up-regulated levels of PTEN and FOXO1.

关 键 词：多发性骨髓瘤 微小RNA-21 细胞株U266 PTEN FOXO1 

分 类 号：Q522[生物学—生物化学] R730.23[医药卫生—肿瘤]