摘要
目的:研究氧化低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVECs)凋亡过程中micro-RNA(miRNA)表达谱的变化,对差异miRNA调控的靶基因进行初步预测。方法:建立体外ox-LDL诱导HUVECs凋亡模型,采用miRNA芯片技术筛选表达变化显著的miRNA,实时荧光定量PCR验证结果,并对差异miRNA调控的靶基因进行生物信息学分析。结果:miRNA芯片检测发现,在ox-LDL诱导HUVECs凋亡过程中有4个表达上调和11个表达下调的miRNA。实时荧光定量PCR对其中2个上调(hsa-miR-142-3p、hsa-miR-365)和2个下调(hsa-miR-590-5p、hsa-miR-33a)miRNA的验证结果与芯片检测所示有较好的一致性。生物信息学分析发现,差异miRNA调控的靶基因与细胞增殖、凋亡、代谢及原癌基因表达等生物学功能相关。结论:经ox-LDL诱导凋亡的HUVECs其miRNA表达谱发生明显改变,提示miRNA可能参与内皮细胞功能障碍和动脉粥样硬化的发生。
Objective： To observe the expression of microRNA （miRNA） in human umbilical vein endothe- lial cells （HUVECs） induced by oxidized low-density lipoprotein （ox-LDL） and to make an initial prediction of target genes regulated by differentially expressed miRNAs. Methods： HUVECs were treated by ox-LDL to estab- lish apoptotic model in vitro, miRNA microarrays were employed to detect the expression profile ofmiRNA in apoptotic HUVECs, and the microarray results were validated by quantitative real-time PCR. The miRNA- targeted genes were predicted by online-available soft-ware. The gene ontology （GO） database and KEGG path- way database were used to determine the functions of these target genes. Results： With the use ofa microarray, we found that ox-LDL up-regulated four miRNAs （hsa-miR-142-3p, hsa-miR-365, hsa-let-7c, hsa-miR-1207-3p） as well as down-regulated eleven miRNAs （hsa-miR-32, hsa-miR-589, hsa-miR-18b, hsa-miR-429, hsa-miR-155, hsa-miR-590-5p, hsa-miR-1197, hsa-miR-222, hsa-miR-374a, hsa-miR-33a, hsa-miR-375） in HUVECs. The expression levels of hsa-miR-142-3p, hsa-miR-365, hsa-miR-590-5p and hsa-miR-33a were validated in accordance with the results of real-time PCR （qRT-PCR）. The bioinformatics analysis indicated that the potential target genes of these aberrantly expressed miRNAs participate in the regu- lation of cell proliferation, apoptosis, metabolism and oncology gene expression. Conclusion： miRNA expres- sion profile in apoptotic HUVECs induced by ox-LDL has significantly changes, which may contribute to en- dothelial dysfunction and development of atherosclerosis.
出处
《神经损伤与功能重建》
2013年第2期80-87,共8页
Neural Injury and Functional Reconstruction
基金
国家自然科学基金(No.81070962)
