Institute of Neurosurgery, Catholic University, Rome, Italy.
J Biol Regul Homeost Agents. 2010 Apr-Jun;24(2):185-95.
Few studies have evaluated the over or the underexpression of genes directly in samples of aneurysmal wall and extracranial pericranial vascular tissue to investigate the genetic influence in formation and rupture of intracranial aneurysms. We present the results obtained using the DNA microarray technique analysis on sample tissues collected during surgery. We collected and analyzed 12 aneurismal and 9 peripheral arteries (superficial temporal (STA) and middle meningeal artery (MMA) specimens from ruptured aneurysm group patients (13 cases), 10 aneurismal and 12 STA and MMA samples from unruptured aneurysm group patients (14 cases) and 5 STA and MMA artery specimens from control group patients (4 cases). Total RNA was isolated from samples and subjected to cDNA microarray analysis with the use of the human genome U133A GeneChip oligonucleotide microarray (Affymetrix, Santa Clara, CA), which allows to analyze a total number of 14,500 genes in the same time. For genes of interest, real-time RT-PCR was performed to confirm their expression level. Total RNA was isolated from samples and subjected to DNA microarray analysis with the use of the human genome U133A GeneChip oligonucleotide microarray, which allows to analyze a total number of 14,500 genes at the same time. For genes of interest, real-time RT-PCR was performed to confirm their expression level. Regarding ruptured aneurysms, genes were identified showing differential expressions (overexpressed or downregulated) pertaining to specific pathways, particularly those for the structural proteins of the extracellular matrix, members of matrix metalloproteinase (MMP) family (which resulted as being overexpressed) and genes involved in apoptotic phenomena. Particularly, real-time RT-PCR analysis confirmed the upregulation of MMP-2, MMP-9 and pro-apoptotic genes, such as Fas, Bax and Bid, and the downregulation of anti-apoptotic genes, such as Bcl-X(L) and Bcl-2. In a compared analyses of ruptured vs unruptured aneurysms, a different expression was also detected regarding gene coding the tissue inhibitor of matrix metalloproteinases 3 (TIMP-3), which appeared markedly downregulated in unruptured aneurysms, where its expression in unruptured aneurysms was similar to that observed in controls. Another gene differently expressed is nitric oxide synthase (iNOS), which appeared overexpressed in ruptured aneurysms when compared to unruptured aneurysms. Our study is the first, to our knowledge, that compares gene expression profiles (genoma-wide) in intracranial aneurysms. The results of our study suggest that the inhibitor of the metalloproteinase, the pathway of nitric oxide and the apoptotic process play a key-role in reducing the resistance of the arterial wall, that can result in formation and rupture of the intracranial aneurysms.
很少有研究直接在动脉瘤壁和颅外血管组织的样本中评估基因的过度或不足表达，以研究遗传因素在颅内动脉瘤的形成和破裂中的作用。我们展示了使用 DNA 微阵列技术在手术中收集的样本组织上进行分析所获得的结果。我们收集并分析了来自破裂性动脉瘤组患者（13 例）的 12 个动脉瘤和 9 个外周动脉（颞浅动脉 [STA] 和脑膜中动脉 [MMA]）样本，来自未破裂性动脉瘤组患者（14 例）的 10 个动脉瘤和 12 个 STA 和 MMA 样本，以及来自对照组患者（4 例）的 5 个 STA 和 MMA 动脉样本。从样本中分离总 RNA，并使用人类基因组 U133A GeneChip 寡核苷酸微阵列（Affymetrix，Santa Clara，CA）进行 cDNA 微阵列分析，该技术可同时分析总共 14500 个基因。对于感兴趣的基因，进行实时 RT-PCR 以确认其表达水平。从样本中分离总 RNA，并使用人类基因组 U133A GeneChip 寡核苷酸微阵列进行 DNA 微阵列分析，该技术可同时分析总共 14500 个基因。对于感兴趣的基因，进行实时 RT-PCR 以确认其表达水平。关于破裂性动脉瘤，确定了显示出差异表达（过度表达或下调）的基因，这些基因涉及特定途径，特别是细胞外基质的结构蛋白、基质金属蛋白酶（MMP）家族的成员（结果显示过度表达）以及涉及凋亡现象的基因。特别是，实时 RT-PCR 分析证实了 MMP-2、MMP-9 和促凋亡基因（如 Fas、Bax 和 Bid）的上调，以及抗凋亡基因（如 Bcl-X（L）和 Bcl-2）的下调。在破裂性动脉瘤与未破裂性动脉瘤的比较分析中，还检测到基因编码组织金属蛋白酶抑制剂 3（TIMP-3）的表达不同，TIMP-3 在未破裂性动脉瘤中的表达明显下调，其在未破裂性动脉瘤中的表达与对照组相似。另一个表达不同的基因是一氧化氮合酶（iNOS），其在破裂性动脉瘤中的表达高于未破裂性动脉瘤。我们的研究是第一个，据我们所知，比较颅内动脉瘤的基因表达谱（基因组范围）。我们的研究结果表明，金属蛋白酶抑制剂、一氧化氮途径和凋亡过程在降低动脉壁阻力方面发挥着关键作用，这可能导致颅内动脉瘤的形成和破裂。
