Microarray identifies ADAM family members as key responders to TGF-beta1 in alveolar epithelial cells.
Microarray identifies ADAM family members as key responders to TGF-beta1 in alveolar epithelial cells.
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DOI：
10.1186/1465-9921-7-114
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发表时间：
2006-09-01
影响因子：
5.8
通讯作者：
Doran, Peter P.
中科院分区：
文献类型：
--
作者：
Keating, Dominic T.;Sadlier, Denise M.;Patricelli, Andrea;Smith, Sinead M.;Walls, Dermot;Egan, Jim J.;Doran, Peter P.
特发性肺纤维化（IPF）的分子机制仍然难以捉摸。转化生长因子β 1（TGF-β1）是肺纤维化发展中的关键效应细胞因子。我们使用微阵列和计算生物学策略来鉴定在肺泡上皮细胞（A549）中响应于TGF-β1、IL-4和IL-13以及Epstein巴尔病毒而表达显著改变的基因。在连续的时间点将A549细胞暴露于10 ng/ml TGF-β1、IL-4和IL-13。总RNA用于与Affyssin人基因组U133 A微阵列杂交。对每个体外时间点进行一式两份研究，并计算平均RMA值。将每个时间点的表达数据与对照进行比较，并采用0.6或更大的信号对数比来鉴定显著的差异调节。使用标准化RMA值和无监督平均连锁分层聚类分析，通过Onto-Compare和Gene-Ontology（GO）数据库管理312种细胞外基质（ECM）蛋白或基质周转调节剂的列表，用于ECM相关基因的诱饵聚类分析。使用本体分类集中的聚类分析对数据集进行询问，揭示了一大群细胞外基质相关基因的协调差异表达。在这一组中，ADAM（含去整合素和金属蛋白酶结构域）基因家族的成员差异表达。在EB病毒感染的A549细胞以及IL-13和IL-4刺激的细胞中也发现了ADAM基因的表达。我们使用siRNA和胶原测定来探测ADAM 19和ADAMTS 9的病理基因组活性（激活和功能活性）。这些基因的敲除导致暴露于TGF-β1的A549细胞中胶原蛋白的产生减少，表明这些分子在IPF中ECM积累中的潜在作用。
The molecular mechanisms of Idiopathic Pulmonary Fibrosis (IPF) remain elusive. Transforming Growth Factor beta 1(TGF-β1) is a key effector cytokine in the development of lung fibrosis. We used microarray and computational biology strategies to identify genes whose expression is significantly altered in alveolar epithelial cells (A549) in response to TGF-β1, IL-4 and IL-13 and Epstein Barr virus. A549 cells were exposed to 10 ng/ml TGF-β1, IL-4 and IL-13 at serial time points. Total RNA was used for hybridisation to Affymetrix Human Genome U133A microarrays. Each in vitro time-point was studied in duplicate and an average RMA value computed. Expression data for each time point was compared to control and a signal log ratio of 0.6 or greater taken to identify significant differential regulation. Using normalised RMA values and unsupervised Average Linkage Hierarchical Cluster Analysis, a list of 312 extracellular matrix (ECM) proteins or modulators of matrix turnover was curated via Onto-Compare and Gene-Ontology (GO) databases for baited cluster analysis of ECM associated genes. Interrogation of the dataset using ontological classification focused cluster analysis revealed coordinate differential expression of a large cohort of extracellular matrix associated genes. Of this grouping members of the ADAM (A disintegrin and Metalloproteinase domain containing) family of genes were differentially expressed. ADAM gene expression was also identified in EBV infected A549 cells as well as IL-13 and IL-4 stimulated cells. We probed pathologenomic activities (activation and functional activity) of ADAM19 and ADAMTS9 using siRNA and collagen assays. Knockdown of these genes resulted in diminished production of collagen in A549 cells exposed to TGF-β1, suggesting a potential role for these molecules in ECM accumulation in IPF.
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影响因子：
82.9
作者：
Fink, L;Seeger, W;Bohle, RM
通讯作者：
Bohle, RM
DOI：
10.1084/jem.194.6.809
发表时间：
2001-09-17
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Lee CG;Homer RJ;Zhu Z;Lanone S;Wang X;Koteliansky V;Shipley JM;Gotwals P;Noble P;Chen Q;Senior RM;Elias JA
通讯作者：
Elias JA
影响因子：
14.9
作者：
Draghici, S;Khatri, P;Tainsky, MA
通讯作者：
Tainsky, MA
DOI：
10.1016/s0167-4781(03)00047-2
发表时间：
2003-04-15
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION
影响因子：
--
作者：
Bevitt, DJ;Mohamed, J;McKie, N
通讯作者：
McKie, N
影响因子：
5.5
作者：
Letterio, JJ;Roberts, AB
通讯作者：
Roberts, AB