ABSTRACT
Background: Metallothioneins (MTs) gene polymorphisms have been associated with the ability of free radicalscavenging and detoxification of heavy metals leading to cancer development. Our aim was to revisit, in a Brazil-ian population, single-nucleotide polymorphisms (SNPs) of the MT gene family previously associated with oralsquamous cell carcinoma (OSCC).Material and Methods: A case-control investigation with 28 OSCC patients and 45 controls was conducted, usingconventional risk factors (tobacco use and alcohol consumption) as covariates. SNPs genotyping for rs8052334(MT1B), rs964372 (MT1B), and rs1610216 (MT2A) was performed by PCR-RFLP, and SNPs for rs11076161(MT1A) were analyzed by TaqMan assay.Results: The only SNP associated with increased risk for OSCC was the MT-1A AA genotype (OR = 4.7; p = 0.01).We have also evidenced for the first time a significant linkage disequilibrium between the SNPs of MT-2A andMT-1A in this population with the highest frequency (30%) of the unfavorable haplotype G/A/C/T (rs1610216 /rs11076161 / rs964372 / rs8052334) of MT gene polymorphisms (OR = 6.2; p = 0.04). Interestingly, after removingthe effects of conventional risk factors, we have uncovered the significance of the AA genotype of the rs11076161with increased odds of 19-fold higher towards OSCC development.Conclusions: This is the first demonstration that a significant linkage disequilibrium among gene polymor-phisms of the MT family may affect susceptibility to oral cancer, which is conditioned by the G/A/C/T haplotype(rs1610216/rs11076161/rs964372/ rs8052334) and the MT-1A gene polymorphism has a potential clinical utility forthe OSCC risk assessment.(AU)
Subject(s)
Humans , Male , Female , Metallothionein , Polymorphism, Genetic , Head and Neck Neoplasms , Carcinoma, Squamous Cell , Squamous Cell Carcinoma of Head and Neck , Mouth Neoplasms/genetics , Brazil , Case-Control Studies , Risk Factors , Oral HealthABSTRACT
Anaplasmosis is one of the most prevalent tick-borne diseases of cattle caused by Anaplasma marginale. MSP1a surface protein has been shown to be involved in eliciting immunity to infected cattle. Carbon nanotubes (CNTs) has been increasingly highlighted due to their needle like structure, which contain multiple attachment sites for biomolecules and may interact with or cross biological membranes, increasing antigen availability to immune system. Here, we have successfully designed a nanocomplex of a synthetic peptide noncovalently attached to multiwalled CNT (MWCNT). Peptide comprising the core motif of the MSP1a was efficiently adsorb onto the nanoparticle surface. The MWCNT-Am1 nanocomplex exhibited high stability and good dispersibility and in vivo immunization showed high levels of IgG1 and IgG2a, followed by increased expression of pro-inflammatory and anti-inflammatory cytokines. This is a proof-of-concept of a nanovaccine that was able to generate a strong immune response compared to the common antigen-adjuvant vaccine without the nanoparticles.
Subject(s)
Anaplasmosis/immunology , Antigens, Bacterial/immunology , Bacterial Outer Membrane Proteins/chemistry , Bacterial Outer Membrane Proteins/immunology , Nanotubes, Carbon/chemistry , Th1 Cells/immunology , Th2 Cells/immunology , Anaplasma/immunology , Anaplasma/pathogenicity , Anaplasmosis/prevention & control , Animals , Antigens, Bacterial/chemistry , Cell Survival/drug effects , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Female , Mice , Mice, Inbred BALB C , Polymerase Chain ReactionABSTRACT
Patients with oral squamous cell carcinoma (OSCC) present significant alterations in their saliva proteome. We have used the shotgun Phage Display (PD) technology to identify candidate proteins that were upregulated in saliva of OSCC by selecting ligands to salivary proteins from a single-chain variable fragment (scFv) PD combinatorial library. After two selection cycles, the highly reactive clone scFv-D09 was able to distinguish saliva of OSCC patients from healthy subjects by enzyme-linked immunosorbent assay (ELISA) with sensitivity and specificity of 96.67%. Additionally, the scFv-D09 clone presented a positive immunostaining for invasive malignant epithelial cells in the connective tissue, keratin pearls in the OSCC, and ducts of salivary glands. We have further identified the target protein as the tropomyosin alpha-4 chain (TPM4) by two-dimensional polyacrylamide gel electrophoresis and mass spectrometry, and its binding to the scFV-D09 was demonstrated by bioinformatics. Briefly, we have identified TPM4 as upregulated salivary protein in patients with OSCC, which plays a central role in stabilizing cytoskeleton actin filaments, probably linked with tumor tissue remodeling. Long-term longitudinal studies are needed to validate TPM4 as a potential marker of a malignant process.
Subject(s)
Carcinoma, Squamous Cell/genetics , Mouth Neoplasms/genetics , Neoplasm Proteins/genetics , Peptide Library , Single-Chain Antibodies/chemistry , Tropomyosin/genetics , Adult , Aged , Aged, 80 and over , Amino Acid Sequence , Carcinoma, Squamous Cell/diagnosis , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/pathology , Computational Biology/methods , Connective Tissue/metabolism , Connective Tissue/pathology , Enzyme-Linked Immunosorbent Assay , Epithelial Cells/metabolism , Epithelial Cells/pathology , Female , Gene Expression , Humans , Male , Middle Aged , Models, Molecular , Mouth Neoplasms/diagnosis , Mouth Neoplasms/metabolism , Mouth Neoplasms/pathology , Neoplasm Proteins/metabolism , Protein Structure, Secondary , Saliva/chemistry , Salivary Glands/metabolism , Salivary Glands/pathology , Sensitivity and Specificity , Tropomyosin/metabolism , Up-RegulationABSTRACT
Properties like high interfacial area with cellular membranes, unique ability to incorporate multiple functionalization, as well as compatibility and transport in biological fluids make carbon nanotubes (CNTs) useful for a variety of therapeutic and drug-delivery applications. Here we used a totally synthetic hybrid supramolecule as an anticancer vaccine formulation. This complex structure comprises CNTs as delivery system for the Cancer Testis Antigen named NY-ESO-1, allied to a synthetic Toll-Like Receptor agonist. The CNT constructs were rapidly internalized into dendritic cells, both in vitro and in vivo, and served as an intracellular antigen depot. This property favored the induction of strong CD4(+) T as well as CD8(+) T cell-mediated immune responses against the NY-ESO-1. Importantly, the vaccination significantly delayed the tumor development and prolonged the mice survival, highlighting the potential application of CNTs as a vaccine delivery system to provide superior immunogenicity and strong protection against cancer.
Subject(s)
CD8-Positive T-Lymphocytes/cytology , Cancer Vaccines/chemistry , Nanotechnology/methods , Nanotubes, Carbon/chemistry , Neoplasms/prevention & control , Neoplasms/therapy , Animals , Anticarcinogenic Agents/chemistry , Antigens/chemistry , Antigens, Neoplasm/chemistry , CD4-Positive T-Lymphocytes/cytology , Calibration , Cell Proliferation , CpG Islands , Dendritic Cells/drug effects , Female , Flow Cytometry , Humans , Lymphocytes/cytology , Membrane Proteins/chemistry , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Microscopy, Electron, Transmission , Neoplasms/metabolism , Oxygen/chemistry , Peptides/chemistry , Spectrum Analysis, RamanABSTRACT
Several studies have been suggesting annexin A1 protein as an active player in tumorigenesis of many organs. Nevertheless, its tumor biomarker role has been mainly studied in tissues by immunohistochemistry or cell culture. Hence, in this investigation, the peripheral blood from 27 oral squamous cell carcinoma (OSCC) patients and 25 negative control individuals were examined by quantitative real-time PCR. Down-regulated ANXA1 expression at mRNA level was observed in OSCC samples (p=0.026). Significantly diminished mRNA levels correlated to age, sex and the anatomical site of the tumor lesion were observed. Moreover, the ROC curve analysis revealed the performance of ANXA1 expression as a suitable biomarker for patients with oral cavity cancer, especially those with 60years of age or older and/or women. For the first time, ANXA1 mRNA is revealed as blood-based biomarker, and its adoption for complementary non-invasive diagnosis of OSCC is suggested. These results suggest that, beyond the anti-inflammatory function, annexin A1 may also play a tumor suppressor role in peripheral blood cells, such as leukocytes.
