Mukhopadhyay P, Singh S, Greene R M, Pisano M M
Department of Molecular Cellular and Craniofacial Biology, University of Louisville Birth Defects Center, ULSD, University of Louisville, KY 40292, USA.
Orthod Craniofac Res. 2006 May;9(2):93-110. doi: 10.1111/j.1601-6343.2006.00356.x.
To determine the differences in gene expression between control-, bone morphogenetic protein (BMP)2- and BMP4-treated murine embryonic maxillary mesenchymal (MEMM) cells.
Transcript profiles of BMP2-, BMP4- and vehicle-treated MEMM cells were compared utilizing the murine high-density GeneChip arrays from Affymetrix. The raw chip data (probe intensities) were pre-processed using robust multichip averaging with GC-content background correction and further normalized with GeneSpring v7.2 software. Cluster analysis of the microarray data was performed with the GeneSpring software. Changes in the gene expression were verified by TaqMan quantitative real-time PCR.
Expression of approximately 50% of the 45 101 genes and expressed sequence tags examined in this study were detected in BMP2-, BMP4- and vehicle-treated MEMM cells and that of several hundred genes was significantly altered (up or downregulated) in these cells in response to BMP2 and BMP4. Expression profiles of each of the 26 mRNAs tested by TaqMan quantitative real-time PCR were found to be consistent with the microarray data. Genes whose expression was modulated following BMP2 or BMP4 treatment, could be broadly classified based on the functions of the encoded proteins such as the growth factors and signaling molecules, transcription factors, and proteins involved in epithelial-mesenchymal interactions, extracellular matrix synthesis, cell adhesion, proliferation, differentiation, and apoptosis.
Utilization of the Affymetrix GeneChip microarray technology has enabled us to delineate a detailed transcriptional map of BMP2 and BMP4 responsiveness in embryonic maxillary mesenchymal cells and offers revealing insights into crucial molecular regulatory mechanisms employed by these two growth factors in orchestrating embryonic orofacial cellular responses.
确定对照处理、骨形态发生蛋白（BMP）2处理和BMP4处理的小鼠胚胎上颌间充质（MEMM）细胞之间的基因表达差异。
利用Affymetrix公司的小鼠高密度基因芯片阵列比较BMP2、BMP4和载体处理的MEMM细胞的转录谱。原始芯片数据（探针强度）使用含GC含量背景校正的稳健多芯片平均法进行预处理，并使用GeneSpring v7.2软件进一步标准化。用GeneSpring软件对微阵列数据进行聚类分析。通过TaqMan定量实时PCR验证基因表达的变化。
在本研究中检测的45101个基因和表达序列标签中，约50%在BMP2、BMP4和载体处理的MEMM细胞中表达，数百个基因的表达在这些细胞中因BMP2和BMP4而显著改变（上调或下调）。通过TaqMan定量实时PCR检测的26种mRNA的表达谱均与微阵列数据一致。其表达在BMP2或BMP4处理后受到调节的基因，可根据编码蛋白的功能大致分类，如生长因子和信号分子、转录因子，以及参与上皮-间充质相互作用、细胞外基质合成、细胞黏附、增殖、分化和凋亡的蛋白。
利用Affymetrix基因芯片微阵列技术使我们能够描绘出胚胎上颌间充质细胞中BMP2和BMP4反应性的详细转录图谱，并为这两种生长因子在协调胚胎口面部细胞反应中所采用的关键分子调控机制提供了深刻见解。
