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1.
Arthritis Rheumatol ; 68(4): 901-14, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26841353

ABSTRACT

OBJECTIVE: In the spondyloarthritides (SpA), increased numbers of CD4+ T cells express killer cell immunoglobulin-like receptor 3DL2 (KIR-3DL2). The aim of this study was to determine the factors that induce KIR-3DL2 expression, and to characterize the relationship between HLA-B27 and the phenotype and function of KIR-3DL2-expressing CD4+ T cells in SpA. METHODS: In total, 34 B27+ patients with SpA, 28 age- and sex-matched healthy controls (20 B27- and 8 B27+), and 9 patients with rheumatoid arthritis were studied. KIR-3DL2 expression and other phenotypic characteristics of peripheral blood and synovial fluid CD4+ T cells were studied by flow cytometry, quantitative polymerase chain reaction, and Western blotting. T cell receptor clonality was determined by template-switch anchored reverse transcription-polymerase chain reaction and sequencing analysis. Cytokines were measured by enzyme-linked immunosorbent assay. RESULTS: Cellular activation induced KIR-3DL2 expression on both naive and effector CD4+ T cells. KIR-3DL2 binding to B27+ cells promoted expression of KIR-3DL2, the Th17-specific transcription factor retinoic acid receptor-related orphan nuclear receptor γt, and the antiapoptotic factor B cell lymphoma 2. KIR-3DL2+CD4+ T cells in patients with ankylosing spondylitis were oligoclonal and enriched for markers of T cell activation and for the gut homing receptor CCR9. In the presence of B27+ antigen-presenting cells, KIR-3DL2+CD4+ T cells produced less interleukin-2 (IL-2) but more IL-17. This effect was blocked by HC10, an antibody that inhibits the binding of KIR-3DL2 to B27 heavy chains. CONCLUSION: KIR-3DL2 binding to HLA-B27 licenses Th17 cell differentiation in SpA. These findings raise the therapeutic potential of targeting HLA-B27-KIR-3DL2 interactions for the treatment of B27+ patients with SpA.


Subject(s)
Cell Differentiation/immunology , HLA-B27 Antigen/immunology , Receptors, KIR3DL2/immunology , Spondylarthropathies/immunology , Th17 Cells/immunology , Adult , Aged , Arthritis, Psoriatic/immunology , Arthritis, Reactive/immunology , Arthritis, Rheumatoid/immunology , Blotting, Western , CD4-Positive T-Lymphocytes/immunology , Case-Control Studies , Cytokines/immunology , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , HLA-B27 Antigen/metabolism , Humans , Male , Middle Aged , Receptors, Antigen, T-Cell/genetics , Receptors, KIR3DL2/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Spondylitis, Ankylosing/immunology , T-Lymphocytes/immunology , Transcriptome , Young Adult
2.
PLoS One ; 10(6): e0129063, 2015.
Article in English | MEDLINE | ID: mdl-26098415

ABSTRACT

OBJECTIVE: The leukocyte immunoglobulin-like receptor (LILR) family includes inhibitory and stimulatory members which bind to classical and non-classical HLA-class I. The ligands for many LILR including LILRB5 have not yet been identified. METHODS: We generated C-terminal eGFP and N-terminal FLAG-tagged fusion constructs for monitoring LILR expression. We screened for LILR binding to HLA-class I by tetramer staining of 293T cells transfected with LILRA1, A4, A5 A6 and LILRB2 and LILRB5. We also studied HLA class I tetramer binding to LILRB5 on peripheral monocyte cells. LILRB5 binding to HLA-class I heavy chains was confirmed by co-immunoprecipitation. RESULTS: HLA-B27 (B27) free heavy chain (FHC) dimer but not other HLA-class I stained LILRB5-transfected 293T cells. B27 dimer binding to LILRB5 was blocked with the class I heavy chain antibody HC10 and anti-LILRB5 antisera. B27 dimers also bound to LILRB5 on peripheral monocytes. HLA-B7 and B27 heavy chains co-immunoprecipitated with LILRB5 in transduced B and rat basophil RBL cell lines. CONCLUSIONS: Our findings show that class I free heavy chains are ligands for LILRB5. The unique binding specificity of LILRB5 for HLA-class I heavy chains probably results from differences in the D1 and D2 immunoglobulin-like binding domains which are distinct from other LILR which bind to ß2m-associated HLA-class I.


Subject(s)
Antigens, CD/metabolism , HLA-B27 Antigen/metabolism , HLA-B7 Antigen/metabolism , Receptors, Immunologic/metabolism , Animals , Cell Line , Humans , Protein Binding , Rats
3.
J Immunol ; 194(4): 1591-601, 2015 Feb 15.
Article in English | MEDLINE | ID: mdl-25582852

ABSTRACT

We proposed that the killer cell Ig-like receptor KIR3DL2 binding more strongly to HLA-B27 (B27) ß2-microglobulin free H chain (FHC) dimers than other HLA-class I molecules regulates lymphocyte function in arthritis and infection. We compared the function of B27 FHC dimers with other class I H chains and identified contact residues in KIR3DL2. B27 FHC dimers interacted functionally with KIR3DL2 on NK and reporter cells more strongly than did other class I FHCs. Mutagenesis identified key residues in the D0 and other Ig-like domains that were shared and distinct from KIR3DL1 for KIR3DL2 binding to B27 and other class I FHCs. We modeled B27 dimer binding to KIR3DL2 and compared experimental mutagenesis data with computational "hot spot" predictions. Modeling predicts that the stronger binding of B27 dimers to KIR3DL2 is mediated by nonsymmetrical complementary contacts of the D0 and D1 domains with the α1, α2, and α3 domains of both B27 H chains. In contrast, the D2 domain primarily contacts residues in the α2 domain of one B27 H chain. These findings provide novel insights about the molecular basis of KIR3DL2 binding to B27 and other ligands and suggest an important role for KIR3DL2-B27 interactions in controlling the function of NK cells in B27(+) individuals.


Subject(s)
HLA-B27 Antigen/immunology , Models, Molecular , Protein Multimerization , Receptors, KIR3DL2/metabolism , Cell Line , Flow Cytometry , HLA-B27 Antigen/chemistry , Humans , Immunoprecipitation , Killer Cells, Natural/immunology , Protein Binding/physiology , Protein Structure, Tertiary/physiology , Receptors, KIR3DL2/chemistry
4.
Mol Immunol ; 57(1): 52-8, 2014 Jan.
Article in English | MEDLINE | ID: mdl-23910730

ABSTRACT

HLA-B27 (B27) is strongly associated with the spondyloarthritides. B27 is expressed at the cell surface of antigen presenting cells (APC) both as canonical ß2m-associated and non-canonical ß2m-free heavy chain (FHC) forms which include B27 dimers (termed B272). B27 FHC forms arise in an endosomal compartment from recycling ß2m-associated B27. Formation of cell surface FHC dimers is critically dependent on an unpaired reactive cysteine 67 in the α1 helix of the class I heavy chain. HLA-B27 also form redox-inducible ß2m-associated dimers on exosomes and apoptosing cells. By contrast with cell surface expressed cysteine 67-dependent heavy chain dimers these dimers are dependent on a cytoplasmic cysteine 325 for their formation. HLA-B27 binds to immunoregulatory receptors including members of the Killer cell Immunoglobulin-like (KIR) and Leukocyte Immunoglobulin-like receptor family. B27 FHC bind to different but overlapping sets of these immunoreceptors compared to classical ß2m-associated HLA-B27. B27 FHC bind more strongly to KIR3DL2 and LILRB2 immune receptor than other ß2m-associated HLA-class I ligands. Genetic studies have implicated genes which control production of the important proinflammatory cytokine IL-17 in the pathogenesis of spondyloarthritis. Cell surface HLA-B27 FHC binding to these immune receptors or acting through other mechanisms could impact on the pathogenesis of spondyloarthritis by promoting immune cell production of IL-17. Here we review the literature on these non-canonical forms of HLA-B27 and the immune receptors they bind to and discuss the possible relevance of these interactions to the pathogenesis of spondyloarthropathy.


Subject(s)
HLA-B27 Antigen/chemistry , HLA-B27 Antigen/metabolism , Membrane Glycoproteins/metabolism , Receptors, Immunologic/metabolism , Receptors, KIR3DL2/metabolism , Spondylitis, Ankylosing/immunology , Spondylitis, Ankylosing/metabolism , Antigens, Surface/chemistry , Antigens, Surface/genetics , Antigens, Surface/immunology , HLA-B27 Antigen/genetics , HLA-B27 Antigen/immunology , Humans , Interleukin-17/biosynthesis , Protein Structure, Secondary , Spondylitis, Ankylosing/genetics
5.
Rheumatology (Oxford) ; 52(11): 1952-62, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23804219

ABSTRACT

OBJECTIVES: HLA-B*27:05 is associated with AS whereas HLA-B*27:09 is not associated. We hypothesized that different interactions with KIR immune receptors could contribute to the difference in disease association between HLA-B*27:05 and HLAB*27:09. Thus, the objective of this study was to compare the formation of ß2m-free heavy chain (FHC) including B27 dimers (B272) by HLA-B*27:05 and HLA-B*27:09 and their binding to KIR immunoreceptors. METHODS: We studied the formation of HLA-B*27:05 and HLA-B*27:09 heterotrimers and FHC forms including dimers in vitro and in transfected cells. We investigated HLA-B*27:05 and HLA-B*27:09 binding to KIR3DL1, KIR3DL2 and LILRB2 by FACS staining with class I tetramers and by quantifying interactions with KIR3DL2CD3ε-reporter cells and KIR3DL2-expressing NK cells. We also measured KIR expression on peripheral blood NK and CD4 T cells from 18 HLA-B*27:05 AS patients, 8 HLA-B27 negative and 12 HLA-B*27:05+ and HLA-B*27:09+ healthy controls by FACS staining. RESULTS: HLA-B*27:09 formed less B272 and FHC than HLA-B*27:05. HLA-B*27:05-expressing cells stimulated KIR3DL2CD3ε-reporter T cells more effectively. Cells expressing HLA-B*27:05 promoted KIR3DL2+ NK cell survival more strongly than HLA-B*27:09. HLA-B*27:05 and HLA-B*27:09 dimer tetramers stained KIR3DL1, KIR3DL2 and LILRB2 equivalently. Increased proportions of NK and CD4 T cells expressed KIR3DL2 in HLA-B*27:05+ AS patients compared with HLA-B*27:05+, HLA-B*27:09+ and HLA-B27- healthy controls. CONCLUSION: Differences in the formation of FHC ligands for KIR3DL2 by HLA-B*27:05 and HLA-B*27:09 could contribute to the differential association of these alleles with AS.


Subject(s)
HLA-B27 Antigen/metabolism , Immunoglobulin Heavy Chains/metabolism , Receptors, KIR3DL2/metabolism , Spondylitis, Ankylosing/genetics , Adult , Alleles , CD4-Positive T-Lymphocytes/immunology , Cell Survival/immunology , Cells, Cultured , Coculture Techniques , Female , Genetic Predisposition to Disease , HLA-B27 Antigen/genetics , Humans , Killer Cells, Natural/immunology , Ligands , Male , Middle Aged , Spondylitis, Ankylosing/immunology , Spondylitis, Ankylosing/metabolism , Transfection
6.
J Immunol ; 190(7): 3216-24, 2013 Apr 01.
Article in English | MEDLINE | ID: mdl-23440420

ABSTRACT

The human leukocyte Ag HLA-B27 (B27) is strongly associated with the spondyloarthritides. B27 can be expressed at the cell surface of APC as both classical ß2-microglobulin-associated B27 and B27 free H chain forms (FHC), including disulfide-bonded H chain homodimers (termed B27(2)). B27 FHC forms, but not classical B27, bind to KIR3DL2. HLA-A3, which is not associated with spondyloarthritis (SpA), is also a ligand for KIR3DL2. In this study, we show that B27(2) and B27 FHC bind more strongly to KIR3DL2 than other HLA-class I, including HLA-A3. B27(2) tetramers bound KIR3DL2-transfected cells more strongly than HLA-A3. KIR3DL2Fc bound to HLA-B27-transfected cells more strongly than to cells transfected with other HLA-class I. KIR3DL2Fc pulled down multimeric, dimeric, and monomeric FHC from HLA-B27-expressing cell lines. Binding to B27(2) and B27 FHC stimulated greater KIR3DL2 phosphorylation than HLA-A3. B27(2) and B27 FHC stimulated KIR3DL2CD3ε-transduced T cell IL-2 production to a greater extent than control HLA-class I. KIR3DL2 binding to B27 inhibited NK IFN-γ secretion and promoted greater survival of KIR3DL2(+) CD4 T and NK cells than binding to other HLA-class I. KIR3DL2(+) T cells from B27(+) SpA patients proliferated more in response to Ag presented by syngeneic APC than the same T cell subset from healthy and disease controls. Our results suggest that expansion of KIR3DL2-expressing leukocytes observed in B27(+) SpA may be explained by the stronger interaction of KIR3DL2 with B27 FHC.


Subject(s)
HLA-B27 Antigen/metabolism , Receptors, KIR3DL2/metabolism , Spondylitis, Ankylosing/immunology , Spondylitis, Ankylosing/metabolism , Antigens/immunology , Cell Line , Cell Survival/immunology , Cells, Cultured , HLA-A3 Antigen/immunology , HLA-A3 Antigen/metabolism , HLA-B27 Antigen/chemistry , HLA-B27 Antigen/immunology , HLA-B35 Antigen/immunology , HLA-B35 Antigen/metabolism , HLA-B7 Antigen/immunology , HLA-B7 Antigen/metabolism , Histocompatibility Antigens Class I/immunology , Histocompatibility Antigens Class I/metabolism , Humans , Interferon-gamma/biosynthesis , Killer Cells, Natural/immunology , Killer Cells, Natural/metabolism , Protein Binding , Protein Multimerization , Receptors, KIR3DL2/genetics , Receptors, KIR3DL2/immunology , Spondylitis, Ankylosing/genetics , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolism
7.
Histol Histopathol ; 28(4): 473-80, 2013 04.
Article in English | MEDLINE | ID: mdl-23382057

ABSTRACT

Fibro-osseous lesions of the jaw are poorly understood because of a significant overlap of clinical, radiological and histological features among the various types, though they present distinct patterns of disease progression. An ossifying fibroma is associated with significant cosmetic and functional disturbances, as it shows expansive proliferation. Thus, it is important to establish a specific marker, as well as clearly elucidate its etiology for diagnosis and proper treatment. We previously established immortalized cell lines from human ossifying fibromas of the jaw and found that they highly expressed the receptor for hyaluronan (HA)-mediated motility (RHAMM). In this study, we examined the expression of RHAMM mRNA in 65 fibro-osseous lesions, including ossifying fibroma, fibrous dysplasia and osseous dysplasia, as well as 5 normal jaws, using real-time RT-PCR and immunohistochemistry assays. RHAMM mRNA and protein expression were significantly elevated in the ossifying fibroma specimens. These results suggest that detection of upregulated RHAMM expression in an ossifying fibroma assists with differential diagnosis and has a key role in elucidation of its pathophysiology.


Subject(s)
Biomarkers, Tumor/analysis , Bone Neoplasms/chemistry , Extracellular Matrix Proteins/analysis , Fibroma, Ossifying/chemistry , Hyaluronan Receptors/analysis , Adolescent , Adult , Aged , Aged, 80 and over , Alkaline Phosphatase/analysis , Biomarkers, Tumor/genetics , Bone Neoplasms/genetics , Bone Neoplasms/pathology , Case-Control Studies , Extracellular Matrix Proteins/genetics , Female , Fibroma, Ossifying/genetics , Fibroma, Ossifying/pathology , Humans , Hyaluronan Receptors/genetics , Immunohistochemistry , Integrin-Binding Sialoprotein/analysis , Male , Middle Aged , Osteocalcin/analysis , Prognosis , RNA, Messenger/analysis , Real-Time Polymerase Chain Reaction , Up-Regulation , Young Adult
8.
J Bone Miner Metab ; 30(3): 293-303, 2012 May.
Article in English | MEDLINE | ID: mdl-21947782

ABSTRACT

Receptor for hyaluronan (HA)-mediated motility (RHAMM) was first described as a soluble HA binding protein released by sub-confluent migrating cells. We previously found that RHAMM was highly expressed and plays an important role in proliferation in the human cementifying fibroma (HCF) cell line, which we previously established. HCF is a benign fibro-osseous neoplasm of the jaw and is composed of fibrous tissue containing varying amounts of mineralized material. However, the pathogenesis of HCF is not clear. In this paper, we examined the roles of RHAMM in osteoblastic cells. We generated RHAMM-overexpressing MC3T3-E1 cells and examined the cell proliferation and differentiation of osteoblastic cells. In MC3T3-E1 cells, overexpressing RHAMM was located intracellular and activated ERK1/2. Interestingly, the ERK1/2 activated by RHAMM overexpression promoted cell proliferation and suppressed the differentiation of osteoblastic cells. Our findings strongly suggest that RHAMM may play a key role in the osteoblastic differentiation process. The rupture of balance from differentiation to proliferation induced by RHAMM overexpression may link to the pathogenesis of bone neoplasms such as HCF.


Subject(s)
Cell Differentiation , Extracellular Matrix Proteins/metabolism , Hyaluronan Receptors/metabolism , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Osteoblasts/cytology , Osteoblasts/enzymology , Animals , Antibodies/pharmacology , Cell Cycle/drug effects , Cell Differentiation/drug effects , Cell Line , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Cell Proliferation/drug effects , Extracellular Matrix Proteins/genetics , Gene Expression Regulation/drug effects , Humans , Hyaluronan Receptors/genetics , Hyaluronan Receptors/immunology , Hyaluronic Acid/pharmacology , Mice , Mitogen-Activated Protein Kinase 1/antagonists & inhibitors , Mitogen-Activated Protein Kinase 3/antagonists & inhibitors , Models, Biological , Osteoblasts/drug effects , Phosphorylation/drug effects , Protein Kinase Inhibitors/pharmacology , Protein Transport/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism
10.
Cancer Lett ; 307(2): 227-36, 2011 Aug 28.
Article in English | MEDLINE | ID: mdl-21570764

ABSTRACT

We found a linear correlation between the Prostaglandin E(2) (PGE(2)) amount and the NR4A2 expression in oral squamous cell carcinoma (SCC) tissues through a statistical analysis among 41 clinical cases. In SCC cell lines, PGE(2) receptor (EP) ligation by exogenous PGE(2) promoted the NR4A2 expression in the cAMP/protein kinase A (PKA)-dependent manner. The process required a nature of SCC cell represented by constitutive activated epidermal growth factor receptor (EGFR) family. Targeted inactivation of the EGFRs interfered the PGE(2)-dependent NR4A2 expression. The PGE(2)-dependent NR4A2 induction is essential for the resistance to anti-cancer drug-induced apoptosis especially in SCC cells which showed constitutive EGFRs activity via autocrine epiregulin, a ligand for EGFRs. Conversely, SCC cells which lack epiregulin expression in their nature could gain the ability to promote the NR4A2 expression in response to PGE(2) and attain the resistance to anti-cancer drug-induced apoptosis under the existence of exogenous epiregulin. These findings suggest that susceptibility of SCC to anti-cancer drug could be compromised when PGE(2) was delivered in the microenvironment of SCC cells supported by constitutive EGFR family activities as their nature.


Subject(s)
Antineoplastic Agents/pharmacology , Carcinoma, Squamous Cell/pathology , Dinoprostone/pharmacology , ErbB Receptors/metabolism , Fluorouracil/pharmacology , Nuclear Receptor Subfamily 4, Group A, Member 2/biosynthesis , Base Sequence , Blotting, Western , Carcinoma, Squamous Cell/drug therapy , Cell Line, Tumor , DNA Primers , Humans , Reverse Transcriptase Polymerase Chain Reaction
11.
Lab Invest ; 91(3): 379-91, 2011 Mar.
Article in English | MEDLINE | ID: mdl-20956971

ABSTRACT

We have previously established immortalized cells (HCF) from cementifying fibroma of the jaw bone. Here, we found that the receptor for hyaluronan (HA)-mediated motility (RHAMM) and epiregulin, a ligand for the epidermal growth factor receptor (EGFR), were highly expressed in HCF cells in comparison with osteoblasts by conducting a microarray analysis. The cell growth of HCF cells was significantly decreased by the knockdown of RHAMM using small interfering RNA (siRNA). RHAMM was associated with extracellular signal-regulated kinase (ERK) and essential for ERK phosphorylation. HCF cells had characteristic growth mechanisms in which epiregulin functions in an extracellular autocrine loop. Interestingly, exogenous HA induced the phosphorylation of EGFR, which was mainly dependent on CD44. The results raise the novel idea that the EGFR may activate Raf-MEK-ERK signaling in response to the binding of HA to CD44. Moreover, RHAMM was able to associate with TPX2 in the nucleus and was required for HA-induced activation of the Aurora A kinase. The results suggest that RHAMM has a predominant role in the cell cycle in HCF. Here, we report the new machinery by which RHAMM/ERK interaction induces the proliferative activity of cementifying fibroma cells via a specific signaling pathway through the CD44-EGFR axis.


Subject(s)
Cell Proliferation , ErbB Receptors/metabolism , Extracellular Matrix Proteins/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , Fibroma, Ossifying/metabolism , Hyaluronan Receptors/metabolism , Aurora Kinases , Cell Cycle Proteins/metabolism , Cell Line, Transformed , Cell Nucleus/metabolism , Enzyme Activation , Epidermal Growth Factor/genetics , Epidermal Growth Factor/metabolism , Epiregulin , ErbB Receptors/antagonists & inhibitors , Extracellular Matrix Proteins/genetics , Fibroma, Ossifying/pathology , Gene Expression Profiling , Gene Silencing , Humans , Hyaluronan Receptors/genetics , Hyaluronic Acid/metabolism , Ligands , Microtubule-Associated Proteins/metabolism , Nuclear Proteins/metabolism , Oligonucleotide Array Sequence Analysis , Phosphorylation , Protein Processing, Post-Translational , Protein Serine-Threonine Kinases/metabolism , RNA, Messenger/metabolism , RNA, Small Interfering , Signal Transduction
12.
J Antibiot (Tokyo) ; 63(2): 77-82, 2010 Feb.
Article in English | MEDLINE | ID: mdl-20057513

ABSTRACT

A new ryanodine-binding inhibitor, verticilide, was isolated from the cultured broth of a fungus, Verticillium sp. FKI-1033. It is a 24-membered ring cyclic depsipeptide, its structure being elucidated as cyclo[(2R)-2-hydroxyheptanoyl-N-methyl- L-alanyl](4). Verticilide inhibited ryanodine binding to ryanodine receptors in the cockroach at an IC(50) value of 4.2 microM, whereas inhibition against mouse ryanodine receptors was weak (IC(50)=53.9 microM).


Subject(s)
Depsipeptides/metabolism , Depsipeptides/pharmacology , Ryanodine Receptor Calcium Release Channel/metabolism , Ryanodine/antagonists & inhibitors , Verticillium/metabolism , Animals , Cell Membrane/metabolism , Cockroaches , Depsipeptides/chemistry , Mice , Molecular Biology , Muscle Fibers, Skeletal/metabolism , Protein Binding , Structure-Activity Relationship
13.
Clin Cancer Res ; 14(19): 6097-105, 2008 Oct 01.
Article in English | MEDLINE | ID: mdl-18829488

ABSTRACT

PURPOSE: Head and neck squamous cell carcinoma (HNSCC) shows persistent invasion that frequently leads to local recurrence and distant lymphatic metastasis. However, molecular mechanisms associated with invasion of HNSCC remain poorly understood. We identified IFN-induced transmembrane protein 1 (IFITM1) as a candidate gene for promoting the invasion of HNSCC by comparing the gene expression profiles between parent and a highly invasive clone. Therefore, we examined the role of IFITM1 in the invasion of HNSCC. EXPERIMENTAL DESIGN: IFITM1 expression was examined in HNSCC cell lines and cases by reverse transcription-PCR and immunohistochemistry. IFITM1 overexpressing and knockdown cells were generated, and the invasiveness of these cells was examined by in vitro invasion assay. Gene expression profiling of HNSCC cells overexpressing IFITM1 versus control cells was examined by microarray. RESULTS: HNSCC cells expressed IFITM1 mRNA at higher levels, whereas normal cells did not. By immunohistochemistry, IFITM1 expression was observed in early invasive HNSCC and invasive HNSCC. Interestingly, IFITM1 was expressed at the invasive front of early invasive HNSCC, and higher expression of IFITM1 was found in invasive HNSCC. In fact, IFITM1 overexpression promoted and IFITM1 knockdown suppressed the invasion of HNSCC cells in vitro. Gene expression profiling of HNSCC cells overexpressing IFITM1 versus control cells revealed that several genes, including matrix metalloproteinase, were up-regulated in IFITM1 overexpressing cells. CONCLUSION: Our findings suggest that IFITM1 plays an important role for the invasion at the early stage of HNSCC progression and that IFITM1 can be a therapeutic target for HNSCC.


Subject(s)
Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms/metabolism , Membrane Proteins/metabolism , Neoplasm Invasiveness , Antigens, Differentiation , Base Sequence , Cell Line, Tumor , Disease Progression , Humans , Immunohistochemistry/methods , Lymphatic Metastasis , Molecular Sequence Data , Oligonucleotide Array Sequence Analysis , RNA Interference , Wound Healing
14.
Planta Med ; 73(14): 1478-81, 2007 Nov.
Article in English | MEDLINE | ID: mdl-17948169

ABSTRACT

NADH-fumarate reductase (NFRD) is a key enzyme in many anaerobic helminths. Decursin and decursinol angelate have been isolated from the roots of ANGELICA GIGAS Nakai (Apiaceae) as NFRD inhibitors. They inhibited ASCARIS SUUM NFRD with IC (50) values of 1.1 and 2.7 microM, respectively. Their target is the electron transport enzyme complex I. Since the inhibitory activities of decursin against bovine heart complexes are weak, it is a selective inhibitor of the nematode complex I. In contrast, decursinol angelate moderately inhibits bovine heart complexes II and III. Decursinol inhibits A. SUUM NFRD to a similar extent, but its target is complex II. It also inhibits bovine heart complexes II and III.


Subject(s)
Anthelmintics/pharmacology , Ascaris suum/enzymology , Benzopyrans/pharmacology , Butyrates/pharmacology , Oxidoreductases Acting on CH-CH Group Donors/antagonists & inhibitors , Angelica/chemistry , Animals , Anthelmintics/chemistry , Benzopyrans/chemistry , Butyrates/chemistry , Molecular Structure
16.
J Antibiot (Tokyo) ; 59(12): 785-90, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17323645

ABSTRACT

A new NADH-fumarate reductase inhibitor, verticipyrone, was isolated from the cultured broth of a fungus, Verticillium sp. FKI-1083. The structure was established as (E)-2-methoxy-3,5-dimethyl-6-(3-methyl-2-undecenyl)-4H-pyran-4-one. Verticipyrone exhibited an IC50 value of 0.88 nM against NADH-fumarate reductase of Ascaris suum. Verticipyrone inhibited both Ascaris and bovine heart complex I, and its synthetic analogue, 8,9-dihydro-8-hydroxyverticipyrone, showed good selectivity against Ascaris complex I.


Subject(s)
Alkenes/isolation & purification , Enzyme Inhibitors/isolation & purification , Oxidoreductases Acting on CH-CH Group Donors/antagonists & inhibitors , Pyrones/isolation & purification , Verticillium/metabolism , Alkenes/chemistry , Alkenes/pharmacology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Fermentation , Magnetic Resonance Spectroscopy , Pyrones/chemistry , Pyrones/pharmacology
17.
J Antibiot (Tokyo) ; 58(1): 50-5, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15813180

ABSTRACT

Nafuredin, a delta-lactone antibiotic, is a fungal metabolite showing selective helminth NADH-fumarate reductase inhibition, and whose target had been revealed as complex I. We found that nafuredin is easily converted to nafuredin-gamma by weak alkaline treatment. The structure of nafuredin-gamma was elucidated as a gamma-lactone form of nafuredin with keto-enol tautomerism. Nafuredin-gamma shows similar complex I inhibitory activity as nafuredin, and it also possesses anthelmintic activity in vivo.


Subject(s)
Anthelmintics/pharmacology , Ascaris suum/drug effects , Pyrones/pharmacology , Succinate Dehydrogenase/antagonists & inhibitors , Animals , Anthelmintics/chemistry , Pyrones/chemistry , Structure-Activity Relationship
18.
J Antibiot (Tokyo) ; 58(1): 74-8, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15813185

ABSTRACT

A new antimycin group antibiotic, antimycin A9, was isolated from a cultured broth of Streptomyces sp. K01-0031 together with antimycins A3a, A3b, A4, and A7, and flazin methyl ester. Antimycin A9 is the first antimycin having an aromatic 8-acyl residue. It showed potent nematocidal and insecticidal activities against Caenorhabditis elegans and Artemia salina, respectively. It inhibited bovine heart NADH oxidase at nanomolar level like other known antimycins.


Subject(s)
Anti-Bacterial Agents/isolation & purification , Antimycin A/analogs & derivatives , Antimycin A/isolation & purification , Antinematodal Agents/isolation & purification , Animals , Anti-Bacterial Agents/pharmacology , Antimycin A/pharmacology , Antinematodal Agents/pharmacology , Artemia/drug effects , Caenorhabditis elegans/drug effects , Cattle , Microbial Sensitivity Tests , Multienzyme Complexes/antagonists & inhibitors , NADH, NADPH Oxidoreductases/antagonists & inhibitors , Streptomyces , Structure-Activity Relationship
19.
J Antibiot (Tokyo) ; 56(6): 533-8, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12931862

ABSTRACT

Structures of three novel compounds designated monordens C to E, isolated from the fermentation broth of amidepsine-producing fungus Humicola sp. FO-2942, were elucidated by spectroscopic evidence. Monordens C is 6,7-dihydromonorden A. Monordens D and E lack the epoxide moiety of monordens C and B, respectively.


Subject(s)
Antifungal Agents/chemistry , Antifungal Agents/isolation & purification , Fermentation , Structure-Activity Relationship
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