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(P<0.05);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(P<0.05)。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(P<0.05);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(P<0.05)。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。
Objective:To investigate the role of microRNAs (miRNAs) in mediating intercellular signaling.Methods:Microparticles (MP) from HUVEC and 293T were isolated by sequential centrifugation.THP-1 was co-cultured with microparticles.And then the migration of THP-1 was measured by transwell.Real-time PCR and Western blotting were used to study the related mechanisms.Results:Compared with the microparticles from 293T,MP from HUVEC could promote the migration of monocytes (P < 0.05) and upregulate the expression of CXCR4 mRNA and protein (P < 0.05).MiRNA-126 deficient MP could downregulate the migration of monocytes (P < 0.05) and the expression of CXCR4 mRNA and protein (P <0.05) compared with miRNA-126 abundant MP.Conclusion:Microparticles from HUVEC could promote the migration of monocytes.As carriers,microparticles could mediate intercellular signaling.
出处
《北京大学学报(医学版)》
CAS
CSCD
北大核心
2014年第6期894-898,共5页
Journal of Peking University:Health Sciences
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~~