Gene expression profiling in lung fibroblasts reveals new players in alveolarization
Gene expression profiling in lung fibroblasts reveals new players in alveolarization
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DOI：
10.1152/physiolgenomics.00108.2007
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发表时间：
2007-12-19
影响因子：
4.6
通讯作者：
Bourbon, Jacques R.
中科院分区：
文献类型：
--
作者：
Boucherat, Olivier;Franco-Montoya, Marie-Laure;Bourbon, Jacques R.
目前对肺泡化的分子基础知之甚少。我们使用了微阵列分析策略，以确定新的基因，可能会调节次级分隔过程。在出生后第2、7和21天，即，分别在分隔之前、期间和之后。提取总RNA，并将cRNA与Affyphin大鼠基因组230 2.0微阵列杂交。通过实时PCR确认选择的基因的表达水平。除了已知在肺泡化过程中上调的基因，包括dreplatin、中期因子、Fgfr 3和Fgfr 4，该研究还使我们能够鉴定出两组具有相反谱的显著基因，即，收集在第7天瞬时上调或下调的基因。前一组包括转录因子视黄酸受体（RXR）-γ和同源框（Hox）a2、a4和a5以及参与Wnt信号传导的基因（Wnt5a、Fzd1和Ndp）;后一组包括细胞外基质组分Comp和Opn以及信号分子Slfn4。从胎儿期到成年期的全肺分析证实了肺泡化的特异性变化。两种治疗，逮捕分隔，高氧和地塞米松，抑制在肺泡化过程中上调的基因的表达，并反过来增强在肺泡化过程中弱表达的基因，此后上调。这些基因在次生分隔的可能作用进行了讨论。对新鲜分离的细胞进行基因表达谱分析是一种强有力的方法，可以提供有关肺泡化过程中基因差异调节和可能参与支气管肺发育不良发病机制的途径的新信息。
Little is known about the molecular basis of lung alveolarization. We used a microarray profiling strategy to identify novel genes that may regulate the secondary septation process. Rat lung fibroblasts were extemporaneously isolated on postnatal days 2, 7, and 21, i.e., before, during, and after septation, respectively. Total RNA was extracted, and cRNAs were hybridized to Affymetrix rat genome 230 2.0 microarrays. Expression levels of a selection of genes were confirmed by real-time PCR. In addition to genes already known to be upregulated during alveolarization including drebrin, midkine, Fgfr3, and Fgfr4, the study allowed us to identify two remarkable groups of genes with opposite profiles, i.e., gathering genes either transiently up- or down-regulated on day 7. The former group includes the transcription factors retinoic acid receptor (RXR)-gamma and homeobox (Hox) a2, a4, and a5 and genes involved in Wnt signaling (Wnt5a, Fzd1, and Ndp); the latter group includes the extracellular matrix components Comp and Opn and the signal molecule Slfn4. Profiling in whole lung from fetal life to adulthood confirmed that changes were specific for alveolarization. Two treatments that arrest septation, hyperoxia and dexamethasone, inhibited the expression of genes that are upregulated during alveolarization and conversely enhanced that of genes weakly expressed during alveolarization and upregulated thereafter. The possible roles of these genes in secondary septation are discussed. Gene expression profiling analysis on freshly isolated cells represents a powerful approach to provide new information about differential regulation of genes during alveolarization and pathways potentially involved in the pathogenesis of bronchopulmonary dysplasia.