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-东南大学学报医学版

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

单位：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

分类号：Q522;R730.23

出版年·卷·期（页码）：2018·37·第五期（819-823）

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：qRT-PCR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.

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