ABSTRACT
<p><b>BACKGROUND</b>Cathepsin L (CatL) is a cysteine protease with strong matrix degradation activity that contributes to photoaging. Mannose phosphate-independent sorting pathways mediate ultraviolet A (UVA)-induced alternate trafficking of CatL. Little is known about signaling pathways involved in the regulation of UVA-induced CatL expression and activity. This study aims to investigate whether a single UVA irradiation affects CatL expression and activity and whether mitogen-activated protein kinase (MAPK)/activator protein-1 (AP-1) pathway is involved in the regulation of UVA-induced CatL expression and activity in human dermal fibroblasts (HDFs).</p><p><b>METHODS</b>Primary HDFs were exposed to UVA. Cell proliferation was determined by a cell counting kit. UVA-induced CatL production and activity were studied with quantitative real-time reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and fluorimetric assay in cell lysates collected on three consecutive days after irradiation. Time courses of UVA-activated JNK and p38MAPK signaling were examined by Western blotting. Effects of MAPK inhibitors and knockdown of Jun and Fos on UVA-induced CatL expression and activity were investigated by RT-PCR, Western blotting, and fluorimetric assay. Data were analyzed by one-way analysis of variance.</p><p><b>RESULTS</b>UVA significantly increased CatL gene expression, protein abundance, and enzymatic activity for three consecutive days after irradiation (F = 83.11, 56.14, and 71.19, respectively; all P < 0.05). Further investigation demonstrated phosphorylation of JNK and p38MAPK activated by UVA. Importantly, inactivation of JNK pathway significantly decreased UVA-induced CatL expression and activity, which were not affected by p38MAPK inhibition. Moreover, knockdown of Jun and Fos significantly attenuated basal and UVA-induced CatL expression and activity.</p><p><b>CONCLUSIONS</b>UVA enhances CatL production and activity in HDFs, probably by activating JNK and downstreaming AP-1. These findings provide a new possible molecular approach for antiphotoaging therapy.</p>
Subject(s)
Child , Child, Preschool , Humans , Anthracenes , Pharmacology , Cathepsin L , Metabolism , Cells, Cultured , Enzyme Inhibitors , Pharmacology , Extracellular Signal-Regulated MAP Kinases , Fibroblasts , Cell Biology , Metabolism , Radiation Effects , Imidazoles , Pharmacology , MAP Kinase Signaling System , Radiation Effects , Oncogene Proteins v-fos , Genetics , Metabolism , Proto-Oncogene Proteins c-jun , Genetics , Metabolism , Pyridines , Pharmacology , Skin , Cell Biology , Ultraviolet RaysABSTRACT
This study aimed to identify the differentially expressed genes after silencing of β-catenin in multiple myeloma transduced with β-catenin shRNA. The DNA microarray dataset GSE17385 was downloaded from Gene Expression Omnibus, including 3 samples of MM1.S (human multiple myeloma cell lines) cells transduced with control shRNA and 3 samples of MM1.S cells transduced with β-catenin shRNA. Then the differentially expressed genes (DEGs) were screened by using Limma. Their underlying functions were analyzed by employing Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. Moreover, DEGs annotation was conducted based on the databases of tumor associated genes, tumor suppressed genes and the transcriptional regulation from patterns to profiles. Furthermore, the protein-protein interaction (PPI) relationship was obtained from STRING and the protein-protein interaction network and the functional modules were visualized by Cytoscape. Then, the pathway enrichment for the DEGs in the functional module was performed. A total of 301 DEGs, including 124 up-regulated and 117 down-regulated DEGs, were screened. Functional enrichment showed that CCNB1 and CDK1 were significantly related to the function of cell proliferation. FOS and JUN were related to innate immune response-activating signal transduction. Pathway enrichment analysis indicated that CCNB1 and CDK1 were most significantly enriched in the pathway of cell cycle. Besides, FOS and JUN were significantly enriched in the Toll-like receptor signaling pathway. FOXM1 was identified as a transcription factor. Moreover, there existed interactions among CCNB1, FOXM1 and CDK1 in PPI network. The expression of FOS, JUN, CCNB1, FOXM1 and CDK1 may be affected by β-catenin in multiple myeloma.
Subject(s)
Humans , CDC2 Protein Kinase , Cyclin B1 , Genetics , Cyclin-Dependent Kinases , Genetics , Forkhead Box Protein M1 , Forkhead Transcription Factors , Genetics , Gene Expression Profiling , Methods , Gene Expression Regulation, Neoplastic , Gene Regulatory Networks , Gene Silencing , Multiple Myeloma , Genetics , Oncogene Proteins v-fos , Genetics , Protein Interaction Maps , Proto-Oncogene Proteins c-jun , Genetics , beta Catenin , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the changes in the plasticity of the neurons and astrocytes in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus of rats exposed to a humid and hot environment.</p><p><b>METHODS</b>The rats were subjected to stimulation with a humid and hot environment for 120 min in a climate chamber (dry bulb temperature of 40.0-/+0.5 degrees C with relative humidity of 60-/+5%). During the exposure, the behavioral responses of the rats were observed, and the changes in the expressions of Fos and GFAP in the PVN and SON in response to the exposure evaluated using immunohistochemical ABC methods.</p><p><b>RESULTS</b>Exposure to a humid and hot environment caused restlessness and agitation in the rats, which showed increased respiratory frequency and scratching of the face with the forelimbs. Two rats died after the 120-min exposure. Significantly increased expressions of Fos and GFAP were detected in the PVN and SON following the exposure as compared with the control group.</p><p><b>CONCLUSION</b>The neurons and astrocytes in the PVN and SON both participate in the regulation of responses to exposure to a humid and hot environment.</p>
Subject(s)
Animals , Male , Rats , Astrocytes , Cell Biology , Physiology , Glial Fibrillary Acidic Protein , Hot Temperature , Humidity , Hypothalamus , Cell Biology , Metabolism , Immunohistochemistry , Neuronal Plasticity , Physiology , Neurons , Cell Biology , Physiology , Oncogene Proteins v-fos , Paraventricular Hypothalamic Nucleus , Cell Biology , Metabolism , Random Allocation , Rats, Sprague-Dawley , Supraoptic Nucleus , Cell Biology , MetabolismABSTRACT
In patients who have sustained traumatic brain injury with associated extremity fracture, there is often a clinical perception that the rate of new bone formation around the fracture site increases.(1) An overgrowth of callus is observed and ectopic ossification even occurs in the muscle,(2) but the mechanism remains unclear. Whether this rapidly-formed new bone is fracture callus or a variant of heterotopic ossification, a common complication of traumatic brain injury, is the subject of some debates.(3) It is generally believed that the process of fracture healing is a recapitulation of normal embryonic osteogenesis,(4) i.e. ,a series of changes in the intracellular and extracellular matrix, which start from the injury of cells, blood vessels and bone matrix to a complete reconstruction of the bone.(5) It is a complex process influenced by multi-level and multi-route regulations of the general and local environments in the body, and many growth factors participate in this process, which is the base of bone healing;(6) whatever methods are used to promote bone healing, they are based on accelerating the changes of growth factors.(7) So it is worth making a thorough study on the mechanism, by which traumatic brain injury influences the expression levels of growth factors and consequently affects the speed of bone healing.
Subject(s)
Animals , Humans , Brain , Metabolism , Brain Injuries , Fibroblast Growth Factor 2 , Physiology , Fracture Healing , Gene Expression , Physiology , Oncogene Protein p65(gag-jun) , Metabolism , Oncogene Proteins v-fos , Metabolism , Vascular Endothelial Growth Factor A , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To observe the effect and mechanism of Tianyuan Ketong recipe (TKR) on the pain reaction in formalin induced pain model rats.</p><p><b>METHODS</b>The analgesic effect of TKR was evaluated using pain behavior graded scoring, the c-fos gene expression and P substance contents in superficial lamella of spinal cord dorsal horn in model rats were analyzed by means of immunohistochemical analysis and computer image analysis technique.</p><p><b>RESULTS</b>TKR could markedly inhibit the pain reaction in model rats (P < 0.05), and the pain induced elevation of c-fos expression and P substance contents could also be suppressed (P < 0.05).</p><p><b>CONCLUSION</b>TKR shows definite analgesic effect on formalin induced pain model rats, the reduction of neuron's reaction in spinal cord dorsal horn to afferent noxious stimulation is possibly one of the pathways for its analgesic effect.</p>
Subject(s)
Animals , Female , Male , Rats , Analgesics, Non-Narcotic , Pharmacology , Drugs, Chinese Herbal , Pharmacology , Formaldehyde , Immunohistochemistry , Oncogene Proteins v-fos , Genetics , Pain , Metabolism , Posterior Horn Cells , Metabolism , Rats, Sprague-Dawley , Spinal Cord , Metabolism , Substance P , MetabolismABSTRACT
The expression of the p53 protein (p53) was compared with those of several oncogenes including c-fos (Fos), c-jun (Jun), and epidermal growth factor receptor (EGFR1) using immunohistochemistry in frozen and paraffin-embedded sections of 25 basal cell carcinomas (BCCs) to find out any correlation between p53 and oncogenes in the pathogenesis of human BCC. In normal skin, positive reactions were obtained for EGFR1 and Fos, while p53 and Jun were negative in all cases. In the lesions, EGFR1 was observed in all cases and p53 was positive in 9 of 25 (36%). Fos was expressed in 21 of 25 (84%) and four negative cases were all p53-positive; this negative correlation between p53 and Fos staining was statistically significant (P< 0.01). Jun was detected in 14 of 20 (70%) and no significant relationship was observed between the expression of Jun and Fos or p53. These data suggest the possibility of down regulation of Fos expression by high levels of p53 protein. Further work is necessary to determine the mechanism of this interaction.