ABSTRACT
Cancer develops when molecular pathways that control the fine balance between proliferation, differentiation, autophagy and cell death undergo genetic deregulation. The prospects for further substantial advances in the management of colorectal cancer reside in a systematic genetic and functional dissection of these pathways in tumor cells. In an effort to evaluate the impact of p38 signaling on colorectal cancer cell fate, we treated HT29, Caco2, Hct116, LS174T and SW480 cell lines with the inhibitor SB202190 specific for p38alpha/beta kinases. We report that p38alpha is required for colorectal cancer cell homeostasis as the inhibition of its kinase function by pharmacological blockade or genetic inactivation causes cell cycle arrest, autophagy and cell death in a cell type-specific manner. Deficiency of p38alpha activity induces a tissue-restricted upregulation of the GABARAP gene, an essential component of autophagic vacuoles and autophagosomes, whereas simultaneous inhibition of autophagy significantly increases cell death by triggering apoptosis. These data identify p38alpha as a central mediator of colorectal cancer cell homeostasis and establish a rationale for the evaluation of the pharmacological manipulation of the p38alpha pathway in the treatment of colorectal cancer.
Subject(s)
Apoptosis/physiology , Autophagy/physiology , HT29 Cells/enzymology , HT29 Cells/pathology , Mitogen-Activated Protein Kinase 14/metabolism , Adaptor Proteins, Signal Transducing/metabolism , Animals , Apoptosis/drug effects , Apoptosis Regulatory Proteins , Autophagy/drug effects , Cell Differentiation , Cell Proliferation , Enzyme Inhibitors/pharmacology , Humans , Mice , Microscopy, Electron , Microtubule-Associated Proteins/metabolism , Mitogen-Activated Protein Kinase 14/deficiency , Mitogen-Activated Protein Kinase 14/drug effects , RNA, Small Interfering/physiology , Tumor Cells, Cultured/enzymologyABSTRACT
The protein product of the retinoblastoma (RB) gene is necessary for the completion of the muscle differentiation program and for myogenic basic helix-loop-helix-dependent transcription. In fact, in addition to induction and maintenance of permanent cell cycle withdrawal through negative regulation of E2F-responsive genes involved in proliferation, pRb also plays a positive role in the activation of muscle-specific genes. In pRb-/- myocytes, the expression of late myogenic markers is defective and myoblast fusion into myotubes occurs without irreversible cell cycle exit. This evidence demonstrates only a partial functional redundancy between pRb and its relatives p107 and pRb2/p130, as these pRb-/- multinucleated cells, which display p107 levels higher than normal myotubes, respond to mitogens with cell cycle re-entry and DNA synthesis. At the molecular level, pRb myogenic functions are mediated by cooperation with MyoD, Myocyte enhancer factor 2 (MEF2), High mobility group box protein-1 (HBP1) and histone deacetylase1, affecting chromatin configuration and tissue-specific transcription, and by post-translational modification in response to intracellular signaling cascades.
Subject(s)
Cell Cycle/physiology , Cell Differentiation/physiology , Genes, Retinoblastoma , Muscle, Skeletal/physiology , Retinoblastoma Protein/physiology , Animals , Cell Communication , Humans , Retinoblastoma Protein/genetics , Signal Transduction , Transcription, GeneticABSTRACT
The optical design of an absorption spectrometer for in situ measurements of atmospheric trace gases is reported. The light source is a rapidly tuned and power-stabilized dye-ring laser, which is frequency doubled by an intracavity BBO crystal. The second harmonic and the fundamental are used simultaneously for measurement of OH, SO(2), CH(2)O, and naphthalene in the UV and of NO(2) in the visible. The 1.2-km absorption path is folded within a 6-m White-cell-type multiple-reflection system with an open-path setup. The absorption sensitivity of the spectrometer is better than 1 part in 10(-5) under tropospheric conditions (integration time 1 min., signal-to-noise ratio 1).
