Project description:The transcriptional program in the response of human fibroblasts to serum control group. This study is described more fully in Iyer VR, et al. 1999. Science 283:83-7 Keywords: other
Project description:The temporal program of gene expression during a model physiological response of human cells, the response of fibroblasts to serum, was explored with a complementary DNA microarray representing about 8600 different human genes. Genes could be clustered in groups on the basis of their temporal patterns of expression in this program. Many features of the transcriptional program appeared to be related to the physiology of wound repair suggesting that fibroblasts play a larger and richer role in this complex multicellular response than had previously been appreciated. This study is described more fully in Iyer VR, et al. 1999. Science 283:83-7 Keywords: other
Project description:The transcriptional program in the response of human fibroblasts to serum. Serum was added to fibroblasts in the presence of cycloheximide. This study is described more fully in Iyer VR, et al. 1999. Science 283:83-7 Keywords: time-course
Project description:The transcriptional program in the response of human fibroblasts to serum. Serum was added to fibroblasts, and samples were taken. This study is described more fully in Iyer VR, et al. 1999. Science 283:83-7 Keywords: time-course
Project description:Foreskin fibroblasts CRL 2091 (ATCC) were serum starved for 48 hours, and harvested at the indicated time points after switching to media with 10% FBS essentially as described (Iyer et al., 1999). RNA from all of the sampled time points were pooled as reference RNA to compare with RNA from individual time points as described (Iyer et al., 1999).
Project description:We used microarrays to profile gene expression changes following growth factor stimulation of primary human fibroblasts. We serum starved (0.1% serum) fibroblasts for 48 hrs and restimulated with 10% serum for 0, 2, 4, 6 and 8 hrs. Total RNA was extracted from 2 independent biological replicas for each time point and hybridized to expression arrays.
Project description:Sharing common ErbB/HER receptor signaling pathway, heregulin (HRG) induces differentiation of MCF-7 breast cancer cells while epidermal growth factor (EGF) elicits proliferation. Although the cell fate led by those two ligands was totally different, the gene expression profile in early transcription was unexpectedly qualitatively similar, suggesting that the gene expression in late transcription, not early transcription, may reflect a respect of ligand specificity. In this study, based on the data from time-course microarray of all human genes, we predicted and determined a series of transcription factors which may control HRG-specific timed-late transcription and cellular differentiation of MCF-7 cells. Validation analyses showed that one of activator protein 1 (AP-1) families appeared just after c-Fos expression, another AP-1 family partner, induced expression of another transcription factor through activation of AP-1 complex. Furthermore, expression of this transcription factors caused suppression of extracellular signal-regulated kinase (ERK) phosphorylation which is sustainedly regulated by HRG-initiated ErbB signaling. Overall, our analysis indicated an importance of formation of timed-transcriptional regulatory network and its function to control upstream signaling pathway through negative feedback for cellular differentiation. Experiment Overall Design: MCF7 human breast cancer cells were stimulated by the growth hormone (epidermal growth factor (EGF) or heregulin (HRG)). Control was set as non-treated cells.
Project description:Mutations in the ABC transporter ABCC6 were recently identified as cause of Pseudoxanthoma elasticum (PXE), a rare genetic disorder characterized by progressive mineralization of elastic fibers. We used an untargeted metabolic approach to identify biochemical differences between human dermal fibroblasts from healthy controls and PXE patients in an attempt to find a link between ABCC6 deficiency, cellular metabolic alterations and disease pathogenesis. 358 compounds were identified by mass spectrometry covering lipids, amino acids, peptides, carbohydrates, nucleotides, vitamins and cofactors, xenobiotics and energy metabolites. We found substantial differences in glycerophospholipid composition, leucine dipeptides, and polypeptides as well as alterations in pantothenate and guanine metabolism to be significantly associated with PXE pathogenesis. These findings can be linked to extracellular matrix remodeling and increased oxidative stress, which reflect characteristic hallmarks of PXE. Our study could facilitate a better understanding of biochemical pathways involved in soft tissue mineralization.