18 research outputs found
TCF/β-catenin plays an important role in HCCR-1 oncogene expression
<p>Abstract</p> <p>Background</p> <p>Oncogene <it>HCCR-1 </it>functions as a negative regulator of the p53 and contributes to tumorigenesis of various human tissues. However, it is unknown how <it>HCCR-1 </it>contributes to the cellular and biochemical mechanisms of human tumorigenesis.</p> <p>Results</p> <p>In this study, we showed how the expression of <it>HCCR-1 </it>is modulated. The luciferase activity assay indicated that the <it>HCCR-1 </it>5'-flanking region at positions -166 to +30 plays an important role in <it>HCCR-1 </it>promoter activity. Computational analysis of this region identified two consensus sequences for the T-cell factor (TCF) located at -26 to -4 (Tcf1) and -136 to -114 (Tcf2). Mutation at the Tcf1 site led to a dramatic decrease in promoter activity. Mobility shift assays (EMSA) revealed that nuclear proteins bind to the Tcf1 site, but not to the Tcf2 site. LiCl, Wnt signal activator by GSK-3β inhibition, significantly increased reporter activities in wild-type Tcf1-containing constructs, but were without effect in mutant Tcf1-containing constructs in HEK/293 cells. In addition, endogenous <it>HCCR-1 </it>expression was also increased by treatment with GSK-3β inhibitor, LiCl or AR-A014418 in HEK/293 and K562 cells. Finally, we also observed that the transcription factor, TCF, and its cofactor, β-catenin, bound to the Tcf1 site.</p> <p>Conclusion</p> <p>These findings suggest that the Tcf1 site on the <it>HCCR-1 </it>promoter is a major element regulating <it>HCCR-1 </it>expression and abnormal stimulation of this site may induce various human cancers.</p
The bone morphogenetic protein antagonist gremlin 1 is overexpressed in human cancers and interacts with YWHAH protein
BACKGROUND: Basic studies of oncogenesis have demonstrated that either the elevated production of particular oncogene proteins or the occurrence of qualitative abnormalities in oncogenes can contribute to neoplastic cellular transformation. The purpose of our study was to identify an unique gene that shows cancer-associated expression, and characterizes its function related to human carcinogenesis. METHODS: We used the differential display (DD) RT-PCR method using normal cervical, cervical cancer, metastatic cervical tissues, and cervical cancer cell lines to identify genes overexpressed in cervical cancers and identified gremlin 1 which was overexpressed in cervical cancers. We determined expression levels of gremlin 1 using Northern blot analysis and immunohistochemical study in various types of human normal and cancer tissues. To understand the tumorigenesis pathway of identified gremlin 1 protein, we performed a yeast two-hybrid screen, GST pull down assay, and immunoprecipitation to identify gremlin 1 interacting proteins. RESULTS: DDRT-PCR analysis revealed that gremlin 1 was overexpressed in uterine cervical cancer. We also identified a human gremlin 1 that was overexpressed in various human tumors including carcinomas of the lung, ovary, kidney, breast, colon, pancreas, and sarcoma. PIG-2-transfected HEK 293 cells exhibited growth stimulation and increased telomerase activity. Gremlin 1 interacted with homo sapiens tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, eta polypeptide (14-3-3 eta; YWHAH). YWHAH protein binding site for gremlin 1 was located between residues 61–80 and gremlin 1 binding site for YWHAH was found to be located between residues 1 to 67. CONCLUSION: Gremlin 1 may play an oncogenic role especially in carcinomas of the uterine cervix, lung, ovary, kidney, breast, colon, pancreas, and sarcoma. Over-expressed gremlin 1 functions by interaction with YWHAH. Therefore, Gremlin 1 and its binding protein YWHAH could be good targets for developing diagnostic and therapeutic strategies against human cancers
Proteomic analysis reveals KRIT1 as a modulator for the antioxidant effects of valproic acid in human bone-marrow mesenchymal stromal cells
<div><p></p><p>Valproic acid (VPA) protects human bone marrow-mesenchymal stromal cells (hBM-MSCs) against oxidative stress and improves their migratory ability through increasing the secretion of trophic factors. This suggests that VPA may be an excellent candidate for improving stem cell function. However, the molecular mechanisms of VPA in BM-MSCs are not known. In this study, we used a proteomic approach to investigate VPA-associated targets under oxidative stress conditions. Krev/Rap1 interaction Trapped-1 (KRIT1), a modulator for the homeostasis of intracellular reactive oxygen species (ROS), was identified as a target protein by two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) analyses. The up-regulation of KRIT1 and its target proteins (SOD2 and FoxO1) with VPA treatment of hBM-MSCs was revealed by qPCR and immunoblot analysis. Damage from oxidative stress was reduced in VPA-pretreated BM-MSCs, which was also confirmed by qPCR and immunoblot analysis. In addition, increased in intracellular ROS by H<sub>2</sub>O<sub>2</sub> were also reduced by VPA pretreatment in BM-MSCs. This suggests that VPA reduces intracellular ROS level by the modulation of KRIT1 and its correlated proteins, FoxO1, SOD2, and cyclin D1. Thus, this study is the first to provide evidence that VPA modulates KRIT1 and intracellular ROS in BM-MSCs.</p></div
The bone morphogenetic protein antagonist gremlin 1 is overexpressed in human cancers and interacts with YWHAH protein-4
<p><b>Copyright information:</b></p><p>Taken from "The bone morphogenetic protein antagonist gremlin 1 is overexpressed in human cancers and interacts with YWHAH protein"</p><p>BMC Cancer 2006;6():74-74.</p><p>Published online 18 Mar 2006</p><p>PMCID:PMC1459871.</p><p>Copyright © 2006 Namkoong et al; licensee BioMed Central Ltd.</p>WHAHp1–60 and YWHAHp81–247 were prepared using YWHAH cDNA as a template DNA. The GST pull-down assay showed that Three GST-YWHAH constructs, Full-YWHAHp1–247, YWHAHp1–100 and YWHAHp1–80 bind YWHAH, but not YWHAHp1–60 and YWHAHp60–247. The YWHAH binding site for PIG-2 was delineated to be residues 61–80 (black box). () Mapping of YWHAH binding domain of PIG-2. Four GST fusion constructs, Full-PIG-2p1-184, PIG-2p1-144, PIG-2p1-100 and PIG-2p1-67 were prepared using PIG-2 cDNA as a template DNA. The GST pull-down assay showed all constructs, The PIG-2 binding site for YWHAH was delineated to be residues 1–67. Gray box is DAN domain. () Schematic diagram shows X-ray crystallography of YWHAH protein. PIG-2 protein binding site was indicated by arrow
Valproic acid promotes neuronal differentiation by induction of neuroprogenitors in human bone-marrow mesenchymal stromal cells
HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-6
<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>d conserved substitutions by gray. (B) Schematic alignment shows the LETM1 domain structure. Analysis using Pfam revealed sequence similarity with the LETM1 domains. Scale is lengths proportional to their molecular masses. The locations of putative LETM domains are indicated by blank boxes. Homo sapiens gi18204589 ref AAH21208; Homo sapiens (HCCR-1) gi13624098 ref AAK34885; Mus musculus gi33416955 ref AAH55685; Drosophila melanogaster gi21626751 ref AAM68317; Drosophila melanogaster gi1749774 ref CAA71125; Molgula oculata, gi308969 ref AAC37181; Caenorhabditis elegans, gi17561658 ref NP_506382; Saccharomyces cerevisiae, gi6324546 ref NP_014615; Saccharomyces cerevisiae, gi1762146 ref AAB70096; Ashbya gossypii gi44980464 ref AAS50397; Ashbya gossypii, gi44980448 ref AAS50381; Neurospora crassa gi32420419 ref XP_330653; Arabidopsis thaliana, gi42562974 ref NP_176732
HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-5
<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>100 nM of staurosporine and 300 nM of actinomycin D, in respectively. The cells were than stained with propidium iodide and analyzed by FACS. The % of apoptotic cells from each analysis were presented on a bar graph
HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-2
<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>ne. Grey boxes indicate the hydrophobic region. TM boxes indicate the locations of putative transmembrane domain. (B) Expression of wild-type or deleted mutants in COS-7 cells. COS-7 cells were transfected with the indicated constructs in the expression vectors. The cells were incubated with MitoTracker and fixed. Fluorescent images of GFP (green), MitoTracker (red) were taken using a confocal microscope. Merged fluorescent images of GFP and MitoTracker are shown. Other conditions are described in Materials and methods
HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-0
<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>d conserved substitutions by gray. (B) Schematic alignment shows the LETM1 domain structure. Analysis using Pfam revealed sequence similarity with the LETM1 domains. Scale is lengths proportional to their molecular masses. The locations of putative LETM domains are indicated by blank boxes. Homo sapiens gi18204589 ref AAH21208; Homo sapiens (HCCR-1) gi13624098 ref AAK34885; Mus musculus gi33416955 ref AAH55685; Drosophila melanogaster gi21626751 ref AAM68317; Drosophila melanogaster gi1749774 ref CAA71125; Molgula oculata, gi308969 ref AAC37181; Caenorhabditis elegans, gi17561658 ref NP_506382; Saccharomyces cerevisiae, gi6324546 ref NP_014615; Saccharomyces cerevisiae, gi1762146 ref AAB70096; Ashbya gossypii gi44980464 ref AAS50397; Ashbya gossypii, gi44980448 ref AAS50381; Neurospora crassa gi32420419 ref XP_330653; Arabidopsis thaliana, gi42562974 ref NP_176732