ABSTRACT
Class A scavenger receptor (SRA), also known as CD204, has been shown to participate in the pathogenesis of atherosclerosis and the pattern recognition of pathogen infection. However, its role in adaptive immune responses has not been well defined. In this study, we report that the lack of SRA/CD204 promotes Toll-like receptor (TLR)4 agonist-augmented tumor-protective immunity, which is associated with enhanced activation of CD8(+) effector T cell and improved inhibition of tumor growth. Dendritic cells (DCs) deficient in SRA/CD204 display more effective immunostimulatory activities upon TLR4 engagement than those from wild-type counterparts. Silencing of SRA/CD204 by RNA interference improves the ability of DCs to prime antigen-specific CD8(+) T cells, suggesting that antigen-presenting cells, for example, DCs, play a major role in SRA/CD204-mediated immune modulation. Our findings reveal a previously unrecognized role for SRA/CD204, a non-TLR pattern recognition receptor, as a physiologic negative regulator of TLR4-mediated immune consequences, which has important clinical implications for development of TLR-targeted immunotherapeutic intervention.
Subject(s)
CD8-Positive T-Lymphocytes/immunology , Down-Regulation/immunology , Lymphocyte Activation/immunology , Scavenger Receptors, Class A/immunology , Signal Transduction/immunology , Toll-Like Receptor 4/immunology , Animals , CD8-Positive T-Lymphocytes/cytology , Cell Line , Cell Proliferation , Dendritic Cells/immunology , Mice , Mice, Inbred C57BL , Mice, Knockout , Neoplasms/immunology , Ovalbumin/immunology , RNA Interference , Scavenger Receptors, Class A/deficiency , Scavenger Receptors, Class A/genetics , Scavenger Receptors, Class A/metabolismABSTRACT
PURPOSE: Peroxiredoxin 1 and 2 are highly homologous members of the Prx (or Prdx) protein family. Prx1 and Prx2 are elevated in several human cancers, and this seems to confer increased treatment resistance and aggressive phenotypes. This study was undertaken to examine the expression profiles of Prx1 and Prx2 in non-small cell lung cancer (NSCLC), and to test their prognostic value in predicting patient survival. EXPERIMENTAL DESIGN: To gain insight into the regulatory mechanisms of Prx1 and Prx2 expression in NSCLC, their respective transcript profiles were examined in NSCLC cell lines from the NCI-60 panel Affymetrix database sets, and the promoter compositions of the two genes were investigated using computer-based multiple sequence alignment analyses. Immunohistochemical analyses of Prx1 and Prx2 were done on a total of 235 NSCLC specimens with stage I through IV disease. The expression profiles of Prx1 and Prx2 in tumor specimens, and their associations with survival, were investigated. RESULTS AND CONCLUSION: The levels of prx1 transcript were higher than those of prx2 in NSCLC cell lines, and the upstream regulatory sequences of the two genes display striking differences. The relative risk of death increased as Prx1 expression levels increased (P = 0.036) in a multivariate Cox model, independent of other clinicopathologic variables associated with survival. No statistically significant correlation was observed between Prx2 and survival. These results suggest that Prx1 may possess unique functions and regulatory mechanisms in NSCLC which are not shared with Prx2, and that Prx1 may serve as a new prognostic biomarker and therapeutic target in NSCLC.
Subject(s)
Carcinoma, Non-Small-Cell Lung/metabolism , Lung Neoplasms/metabolism , Peroxiredoxins/physiology , Base Sequence , Carcinoma, Non-Small-Cell Lung/mortality , Carcinoma, Non-Small-Cell Lung/therapy , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Humans , Immunohistochemistry , Lung Neoplasms/mortality , Lung Neoplasms/therapy , Molecular Sequence Data , Peroxiredoxins/analysis , Peroxiredoxins/antagonists & inhibitors , Peroxiredoxins/genetics , Prognosis , Promoter Regions, Genetic , RNA, Messenger/analysis , Up-RegulationABSTRACT
The major estrogen metabolite 2-methoxyestradiol (2ME) has been shown to target tumor cells without severe side effects and is currently being evaluated in clinical trials for several types of cancer. Despite its promise for use in clinical setting, the mechanism(s) by which 2ME exerts its anti-tumor activity is not clearly defined at this time. Employing organic chemistry tools, we synthesized 2ME analogs with which 2ME affinity column was prepared, enabling us to detect a protein that selectively interacts with 2ME. This 2ME analog will be useful as a probe to identify the biological target(s) of 2ME and study their functions in tumor cells.
Subject(s)
Antineoplastic Agents/pharmacology , Estradiol/analogs & derivatives , 2-Methoxyestradiol , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Cell Line, Tumor , Cell Proliferation/drug effects , Drug Screening Assays, Antitumor , Estradiol/chemical synthesis , Estradiol/chemistry , Estradiol/pharmacology , Humans , Molecular Conformation , Stereoisomerism , Structure-Activity RelationshipABSTRACT
Generation of a monomethylated selenium metabolite is critical for the anticancer activity of selenium. Because of its strong nucleophilicity, the metabolite can react directly with protein thiols to cause redox modification. Here, we report a neural network-based analysis to identify potential selenium targets. A reactive thiol specific reagent, BIAM, was used to monitor thiol proteome changes on 2D gel. We constructed a dynamic model and evaluated the relative importance of proteins mediating the cellular responses to selenium. Information from this study will provide new clues to unravel mechanisms of anticancer action of selenium. High impact selenium targets could also serve as biomarkers to gauge the efficacy of selenium chemoprevention.
Subject(s)
Neural Networks, Computer , Proteome/analysis , Proteomics/methods , Selenium/pharmacology , Sulfhydryl Compounds/analysis , Antineoplastic Agents/metabolism , Antineoplastic Agents/pharmacology , Biotin/analogs & derivatives , Biotin/chemistry , Cell Line, Tumor , Electrophoresis, Gel, Two-Dimensional , Ethylenediamines/chemistry , Humans , Male , Organoselenium Compounds/metabolism , Organoselenium Compounds/pharmacology , Oxidation-Reduction/drug effects , Prostatic Neoplasms/drug therapy , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/pathology , Proteins/analysis , Proteins/chemistry , Proteins/metabolism , Proteome/chemistry , Selenium/metabolism , Spectrometry, Mass, Electrospray Ionization , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Sulfhydryl Compounds/chemistry , Sulfhydryl Compounds/metabolismABSTRACT
We examined hypoxia-induced changes in global thiol proteome profile in human prostate cancer cells using a BIAM-based display method. We analyzed the kinetics of protein thiol modification by using a pattern recognition algorithm, self-organizing maps (SOM) clustering, and identified the BIAM-labeled proteins by MALDI-TOF and ESI-tandem mass spectrometry. We found 99 out of 215 of total BIAM-labeled proteins were affected by hypoxia treatment and, yet, with diverse patterns and kinetics of redox modification. Our study proved that proteomics analysis employing the BIAM-labeling method can provide valuable information pertaining to global changes in the redox status of proteins in response to hypoxia.
Subject(s)
Proteome/analysis , Proteomics/methods , Apoptosis/physiology , Biotin/analogs & derivatives , Biotin/chemistry , Caspases/metabolism , Cell Hypoxia/physiology , Cell Line, Tumor , Cluster Analysis , Electrophoresis, Gel, Two-Dimensional , Ethylenediamines/chemistry , Humans , JNK Mitogen-Activated Protein Kinases/metabolism , Oxidation-Reduction , Poly(ADP-ribose) Polymerases/metabolism , Proteins/analysis , Proteins/chemistry , Proteins/metabolism , Proteome/metabolism , Reactive Oxygen Species/metabolism , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
Small molecules designed to specifically activate or inactivate protein functions have been useful to study biological processes. PROTACS are small molecule chimera which comprise a ligand and a peptide recognition motif for an E3 ligase. These novel reagents exploit the ubiquitin-mediated proteasome degradation pathway to target the ligand-bound protein for intracellular degradation. Here, we report that an estrogen receptor (ER)-targeting PROTACS that causes degradation of ER is able to potently inhibit endothelial cell differentiation in a three-dimensional angiogenic sprouting assay. These findings support the use of ER-targeting PROTACS as probes of angiogenesis.
Subject(s)
Neovascularization, Physiologic , Proteasome Endopeptidase Complex/metabolism , Receptors, Estrogen/metabolism , Ubiquitin/metabolismABSTRACT
Ubiquitin-dependent proteolysis of cellular proteins is one of the major pathways to regulate protein function posttranslationally. Here we demonstrate a potentially general method of degrading any targeted proteins by the ubiquitin-dependent proteolysis in living cells, using small-molecule proteolysis inducer (SMPI).
Subject(s)
Multienzyme Complexes/metabolism , Peptide Fragments/metabolism , Peptide Hydrolases/metabolism , Proteins/metabolism , Ubiquitin/metabolism , Humans , Hypoxia-Inducible Factor 1, alpha Subunit , Multienzyme Complexes/chemistry , Peptide Fragments/chemistry , Peptide Hydrolases/chemistry , Proteins/chemistry , Transcription Factors/metabolism , Tumor Cells, Cultured , Ubiquitin/chemistry , Ubiquitin-Protein Ligases/metabolismABSTRACT
Cavernous hemangiomas are one of the most common benign tumors of the orbit in adults. We report a case of a longstanding retrobulbar hemangioma that was removed successfully through a temporal transconjunctival approach combined with lateral canthotomy. A 45-year-old female patient, with a 15-year history of slowly progressive proptosis and decreased visual acuity of the left eye, had a corrected visual acuity of finger count at 50 cm OS, compared with 1.0 OD. Exophthalmometry by the Nagle's method measured 15 mm OD by 26 mm OS. Magnetic resonance imaging (MRI) revealed a well-encapsulated retrobulbar main mass, 2.3 x 3.0 x 3.7 cm in size along with multiple small satellite nodules that were displacing the optic nerve and globe superiorly. The tumors were removed through a superotemporal transconjunctival approach combined with lateral canthotomy. Pathological examination revealed an intraorbital cavernous hemangioma. The patient was free of visible scars, proptosis and any other noticeable complications at her last follow-up, 6 months after surgery.
