【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。更多还原

【Abstract】 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 qRTPCR. 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.更多还原