Simultaneous analysis of 1176 gene products in normal human aorta and abdominal aortic aneurysms using a membrane-based complementary DNA expression array
Simultaneous analysis of 1176 gene products in normal human aorta and abdominal aortic aneurysms using a membrane-based complementary DNA expression array
复制标题
DOI：
10.1067/mva.2001.113310
复制
发表时间：
2001-07-01
影响因子：
4.3
通讯作者：
Thompson, RW
中科院分区：
文献类型：
--
作者：
Tung, WS;Lee, JK;Thompson, RW
背景：腹主动脉瘤（AAAs）中已经描述了一些基因表达的变化，但这种疾病的分子改变谱尚不清楚。本研究的目的是表征人类AAA组织中大约1000个基因产物的表达，并将AAA组织中表达的基因与正常主动脉中观察到的基因进行比较。材料和方法从腹主动脉壁组织（4条AAAs和4条正常主动脉）中分离总RNA，通过反转录构建阵列特异性的[P-32]标记的互补DNA （cDNA）探针。将cDNA探针与含有1176个cDNA克隆阵列（AtlasArray Human 1.2 I; Clontech, palalalto, california）的尼龙膜杂交，扫描放射自显像以确定每种组织类型的基因表达特征模式。密度分析用于将单个基因的表达标准化到一组管家控制，差异基因表达的信号比至少为2:1。结果：1176个基因中145个（12.3%）在主动脉组织中一致表达。在AAAs和正常主动脉中检测到的101个转录本中，胸腺素β -4含量最高，而44个基因表现出差异表达模式（39个在AAAs中占优势，5个在正常主动脉中占优势）。密度分析证实了AAAs与正常主动脉之间20种基因产物的表达差异，其中髓样细胞核分化抗原（31倍）、组织蛋白酶H（30倍）、血小板衍生生长因子- a（23倍）、载脂蛋白E（13倍）、明胶酶B/基质金属蛋白酶-9（12倍）和白细胞介素-8（11倍）的表达差异最大。只有肌球蛋白轻链激酶（39倍）和β -1整合素（2倍）的基因产物在AAAs中显著减少。因此AAA组织表现出一种独特的基因表达模式，反映慢性炎症、细胞外基质降解、动脉粥样硬化和平滑肌细胞耗损。结论：cDNA表达阵列为确定动脉瘤变性的分子机制提供了一种强有力的新方法。需要进一步的研究来阐明在AAA组织中表现出表达水平改变的单个基因的功能和病理生理意义。
Background: A number of changes in gene expression have been described in abdominal aortic aneurysms (AAAs), but the spectrum of molecular alterations in this disease is un]known. The purpose of this study was to characterize the expression of approximately 1000 gene products in human AAA tissue and to compare the profile of genes expressed in AAAs with that observed in normal aorta.Materials and Methods Total RNA was isolated from abdominal aortic wall tissues (4 AAAs and 4 normal aortas), and array-specific [P-32]-labeled complementary DNA (cDNA) probes were created with reverse transcription. The cDNA probes were hybridized with nylon membranes containing an array of 1176 cDNA clones (AtlasArray Human 1.2 I; Clontech, Pale Alto, Calif), and autoradiographs were scanned to identify the patterns of gene expression characteristic of each tissue type. Densitometric analysis was used to standardize the expression of individual genes to a panel of housekeeping controls, and differential gene expression was defined by a signal ratio of at least 2:1.Results: One hundred forty-five (12.3%) of the 1176 genes were consistently expressed in aortic tissue. Thymosin beta -4 was the most abundant of 101 transcripts detected in both AAAs and normal aorta, whereas 44 genes exhibited differential patterns of expression (39 predominant in AAAs and 5 in normal aorta). Densitometric analysis confirmed differences in expression for 20 of these gene products between AAAs and normal aorta, with the greatest increases seen for myeloid cell nuclear differentiation antigen (31-fold), cathepsin H (30-fold), platelet-derived growth factor-A (23-fold), apolipoprotein E (13-fold), gelatinase B/matrix metalloproteinase-9 (12-fold), and interleukin-8 (11-fold). The only gene products substantially decreased in AAAs were myosin light chain kinase (39-fold) and beta -1 integrin (twofold). AAA tissues thereby exhibited a distinct pattern of gene expression reflecting chronic inflammation, extracellular matrix degradation, atherosclerosis, and smooth muscle cell depletion.Conclusion: cDNA expression arrays provide a powerful new approach to help identify the molecular mechanisms responsible for aneurysmal degeneration. Further studies will be needed to elucidate the functional and pathophysiologic significance of the individual genes that exhibit altered levels of expression in AAA tissue.