Sukhanov Sergiy, Delafontaine Patrick
Tulane University Health Sciences Center, New Orleans, LA 70112, USA.
Proteomics. 2005 Apr;5(5):1274-80. doi: 10.1002/pmic.200400985.
Commercially available high-content Ab380 and extensively validated DLM26 homemade protein microarrays were used to profile the effects of the pro-atherogenic molecule, oxidized low density lipoprotein (OxLDL), on human aortic smooth muscle cells. Protein microarrays detected 298 proteins in cell lysates and 54 of these were differentially regulated. Microarray data were validated by immunoblotting for a selected set of up- and down-regulated proteins. The protein microarray data sets were compared with our recent cDNA microarray-based gene expression results in order to characterize the global effect of OxLDL on smooth muscle cell functions. A group of cell-cell interaction molecules was classified as up-regulated by OxLDL, whereas nucleic acid/protein biosynthesis, structural and humoral response proteins/genes were under-expressed in cells treated by OxLDL. These findings reveal the major pattern of OxLDL-induced effects on the human aortic smooth muscle cells functions and also demonstrate that protein chip-based microarrays could be a useful proteomic tool to profile disease-related states of muscle cells.
使用市售的高含量Ab380和经过广泛验证的DLM26自制蛋白质微阵列，来分析促动脉粥样硬化分子氧化型低密度脂蛋白（OxLDL）对人主动脉平滑肌细胞的影响。蛋白质微阵列检测到细胞裂解物中的298种蛋白质，其中54种蛋白质的表达存在差异调节。通过对一组选定的上调和下调蛋白质进行免疫印迹，验证了微阵列数据。将蛋白质微阵列数据集与我们最近基于cDNA微阵列的基因表达结果进行比较，以表征OxLDL对平滑肌细胞功能的整体影响。一组细胞间相互作用分子被归类为受OxLDL上调，而核酸/蛋白质生物合成、结构和体液反应蛋白/基因在经OxLDL处理的细胞中表达下调。这些发现揭示了OxLDL对人主动脉平滑肌细胞功能诱导作用的主要模式，也证明基于蛋白质芯片的微阵列可能是一种用于分析肌肉细胞疾病相关状态的有用蛋白质组学工具。
