Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from Mus musculus tissues (Heart, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from seven Mus musculus tissues (Heart, Brain, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:The Forkhead Box m1 (Foxm1 or Foxm1b) protein (previously called HFH-11B, Trident, Win, or MPP2) is abundantly expressed in human non-small cell lung cancers where it transcriptionally induces expression of genes essential for proliferation of tumor cells. In this study, we used Rosa26-Foxm1 transgenic mice, in which the Rosa26 promoter drives ubiquitous expression of Foxm1 transgene, to identify new signaling pathways regulated by Foxm1. Lung tumors in Rosa26-Foxm1 mice were induced using the 3-methylcholanthrene (MCA)/ butylated hydroxytoluene (BHT) lung tumor initiation/ promotion protocol. MCA/BHT-treated Rosa26-Foxm1 mice displayed a significant increase in the proliferation of lung tumor cells as well as in the number and size of lung adenomas. Elevated tumor formation in Rosa26-Foxm1 transgenic lungs was associated with persistent pulmonary inflammation, macrophage infiltration and increased expression of Cdc25C phosphatase, cyclin E2, chemokine ligands Cxcl5, Cxcl1 and Ccl3, cathepsins, and Matrix metalloprotease 12, all of which stimulate signaling pathways essential for cellular proliferation, pulmonary inflammation and remodeling. Lung tumors from Rosa26-Foxm1 mice displayed increased expression of Cyclooxygenase-2 (Cox-2), whereas diminished Cox-2 levels were observed in Foxm1-deficient lung tumors from Mx-Cre Foxm1 fl/fl mice. Cell culture experiments with A549 human lung adenocarcinoma cells demonstrated that depletion of Foxm1 by either siRNA transfection or treatment with Foxm1-inhibiting ARF 26-44 peptide significantly reduced Cox-2 expression. Foxm1 protein bound to the –3479/–3461 and –7826/–7795 bp regions of the human Cox-2 promoter, indicating that the human Cox-2 gene is a direct transcriptional target of Foxm1 in lung tumor cells. Keywords: Influence of genetic modification to the tumor development
Project description:The importance of unanchored Ub in innate immunity has been shown only for a limited number of unanchored Ub-interactors. We investigated what additional cellular factors interact with unanchored Ub and whether unanchored Ub plays a broader role in innate immunity. To identify unanchored Ub-interacting factors from murine lungs, we used His-tagged recombinant poly-Ub chains as bait. These chains were mixed with lung tissue lysates and protein complexes were isolated with Ni-NTA beads. Sample elutions were subjected to mass spectrometry (LC-MSMS) analysis.
Project description:MYBL2 is a transcription factor that has either pro-survival or anti-survival functions in a cell-type specific manner. Overexpression of MYBL2 is associated with worse survival of lung adenocarcinoma, but the mechanism by which it regulates transcription has not yet been elucidated. In this study, we found that MYBL2 mainly binds to the promoters of highly expressed genes in lung adenocarcinoma cells using ChIP-seq. Using knock-down and RNA-seq approach, we identified over a thousand of genes deregulated by MYBL2. By integrating ChIP-seq and RNA-seq data, we identified target genes of MYBL2. We revealed that FOXM1 is regulated by MYBL2 in lung adenocarcinoma cells, and FOXM1 binding sites are largely shared with MYBL2 binding sites. We treated lung adenocarcinoma cells with FDI-6, a known FOXM1 inhibitor and investigated the effect of FDI-6 in transcriptional regulation of MYBL2 and FOXM1. We found that CENPA is one of the key genes regulated by MYBL2 and FOXM1, and that it can be inhibited by FDI-6. Our signaling pathway analysis results revealed that MYBL2 and FOXM1 activate cell-cycle genes, suggesting that MYBL2 and FOXM1 act as oncogenic transcription factors in lung adenocarcinoma cells and FDI-6 could be a potential treatment of the disease.