微颗粒携带microRNA-126在细胞间信号传递中的作用

发布时间：2018-04-30 14:32

本文选题：微颗粒 + 微RNAs　；参考：《北京大学学报(医学版)》2014年06期

【摘要】：目的:探讨微颗粒(microparticles,MP)作为载体携带microRNAs(miRNAs)在介导细胞间信号传递中的作用。方法:用梯度离心的方法获得人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)和293T细胞来源的微颗粒,将微颗粒与人急性单核细胞白血病细胞(THP-1细胞)共培养,然后应用transwell迁移实验观察THP-1细胞迁移情况;用实时定量聚合酶链反应(polymerase chain reaction,PCR)及Western blotting实验的方法,分析微颗粒携带miRNAs影响单核细胞迁移的机制。结果:通过transwell实验发现,HUVEC细胞来源的微颗粒(MPHUVEC)组THP-1细胞迁移数明显多于293T细胞来源的微颗粒(MP293T)组,差异有统计学意义(P0.05);MPHUVEC组THP-1细胞CXCR4 mRNA及蛋白表达水平均明显高于MP293T组,差异有统计学意义(P0.05)。HUVEC细胞分别给予lipo2000、lipo2000+miRNA inhibitor和lipo2000+miRNA-126 inhibitor不同处理获得的微颗粒为MPcon、MPneg-con、MPmiR-126 inhibitor,与MPcon组、MPneg-con组相比,MPmiR-126 inhibitor组THP-1细胞迁移数明显减少,差异有统计学意义(P0.05);MPmiR-126 inhibitor组THP-1细胞CXCR4 mRNA及蛋白表达水平显著低于MPcon组、MPneg-con组,差异有统计学意义(P0.05)。结论:HUVEC细胞来源的微颗粒可以携带miRNA-126促进单核细胞迁移,微颗粒作为载体可以携带miRNAs介导细胞间信号传递。
[Abstract]:Aim: to investigate the role of microgranular MPs as a carrier carrying microRN AsmiRNs in intercellular signal transduction. Methods: human umbilical vein endothelial cells and 293T cells derived from human umbilical vein endothelial cells (HUVECs) and 293T cells were obtained by gradient centrifugation and co-cultured with human acute monocytic leukemia cells (THP-1 cells). Then the migration of THP-1 cells was observed by transwell migration assay, and the mechanism of micronucleus carrying miRNAs on monocyte migration was analyzed by real-time quantitative polymerase chain reaction- PCR and Western blotting assay. Results: transwell assay showed that the number of migration of THP-1 cells in the MPHUVEC-derived group was significantly higher than that in the MP293T group, and the expression of CXCR4 mRNA and protein in the THP-1 cells in the P0.05-MPHUVEC group was significantly higher than that in the MP293T group. There were significant differences in the number of THP-1 cell migration between MPcon group and MPneg-con-lated group. The migration of THP-1 cells in the MPmiR-126 inhibitor group was significantly lower than that in the MPcon group compared with the MPcon group. The expression of CXCR4 mRNA and protein in THP-1 cells in P0.05 inhibitor group was significantly lower than that in MPneg-con group (P 0.05). Conclusion the microparticles derived from the cells can carry miRNA-126 to promote monocyte migration, and microparticles can carry miRNAs to mediate intercellular signal transduction.
【作者单位】：北京大学人民医院心脏中心;
【基金】：国家自然科学基金(81270274) “重大新药创制”科技重大专项(2012ZX09303019) 首都卫生发展科研专项项目(2011-4022-03) 北京市自然科学基金(7132225) 北京大学人民医院研究与发展基金(RDB2011-17)资助~~
【分类号】：R329.2

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