Archacki Stephen R, Wang Qing K
Center for Molecular Genetics, The Cleveland Clinic Foundation, OH, USA.
Methods Mol Med. 2006;129:1-13. doi: 10.1385/1-59745-213-0:1.
Microarray analysis is a powerful technique for high-throughput, global transcriptonomic profiling of gene expression. It holds great promise for analyzing the genetic and molecular bases of cardiovascular diseases and various other complex diseases and permits the analysis of thousands of genes simultaneously, both in diseased and nondiseased tissues and/or cell lines. Microarrays or microchips are made by depositing spots of DNA or oligonucleotides representing thousands of genes on a solid support such as a coated glass surface, and can allow the comparison of gene expression patterns in any two samples. Total RNA is isolated from the tissue or cells of interest, converted to cDNA and then cRNA labeled with biotin, and hybridized to the chips. Hybridization signals are then quantified and compared among different samples. We used oligonucleotide microarrays to obtain an unbiased assessment of expression levels of thousands of genes simultaneously in normal and diseased coronary arteries. Fifty-six genes showed differential expression in atherosclerotic coronary artery tissues, and 49 of them represent new linked genes for coronary artery disease. These studies can generate novel hypotheses relating to the pathologies of disease and further studies with animal models, molecular biology, cell biology, and biochemistry will validate these hypotheses and provide novel insights into the pathogenesis of disease.
微阵列分析是一种用于基因表达高通量、全转录组分析的强大技术。它在分析心血管疾病及其他各种复杂疾病的遗传和分子基础方面具有巨大潜力，能够同时对患病和未患病组织及/或细胞系中的数千个基因进行分析。微阵列或微芯片是通过将代表数千个基因的DNA或寡核苷酸点样沉积在诸如涂覆玻璃表面等固体支持物上制成的，可用于比较任意两个样本中的基因表达模式。从感兴趣的组织或细胞中分离出总RNA，将其转化为cDNA，然后用生物素标记cRNA，并与芯片杂交。接着对杂交信号进行定量并在不同样本之间进行比较。我们使用寡核苷酸微阵列同时对正常和患病冠状动脉中数千个基因的表达水平进行了无偏评估。56个基因在动脉粥样硬化冠状动脉组织中表现出差异表达，其中49个代表冠状动脉疾病的新关联基因。这些研究能够产生与疾病病理学相关的新假说，而进一步利用动物模型、分子生物学、细胞生物学和生物化学进行的研究将验证这些假说，并为疾病发病机制提供新的见解。
