mart-1-antigen and Cell-Transformation--Neoplastic

Topics: Adult; Angiomyolipoma; Biomarkers, Tumor; Carcinoma, Hepatocellular; Cell Transformation, Neoplastic; Diagnosis, Differential; Female; gp100 Melanoma Antigen; Humans; Kidney Neoplasms; Liver Neoplasms; MART-1 Antigen; Melanoma-Specific Antigens; Neoplasms, Multiple Primary; Tomography, X-Ray Computed; Tuberous Sclerosis; Young Adult

Anoikis resistance is one of the abilities acquired along tumor progression. This characteristic is associated with metastasis development, since tumorigenic cells must survive independently of cell-matrix interactions in this process. In our laboratory, it was developed a murine melanocyte malignant transformation model associated with a sustained stressful condition. After subjecting melan-a melanocytes to 1, 2, 3 and 4 cycles of anchorage impediment, anoikis resistant cells were established and named 1C, 2C, 3C and 4C, respectively. These cells showed altered morphology and PMA independent cell growth, but were not tumorigenic, corresponding to pre-malignant cells. After limiting dilution of 4C pre-malignant cells, melanoma cell lines with different characteristics were obtained. Previous data from our group showed that increased Timp1 expression correlated with anoikis-resistant phenotype. Timp1 was shown to confer anchorage-independent growth capability to melan-a melanocytes and render melanoma cells more aggressive when injected into mice. However, the mechanisms involved in anoikis regulation by Timp1 in tumorigenic cells are not clear yet.. The β1-integrin and Timp1 expression were evaluated by Western blotting and CD63 protein expression by flow cytometry using specific antibodies. To analyze the interaction among Timp1, CD63 and β1-integrin, immunoprecipitation assays were performed, anoikis resistance capability was evaluated in the presence or not of the PI3-K inhibitors, Wortmannin and LY294002. Relative expression of TIMP1 and CD63 in human metastatic melanoma cells was analyzed by real time PCR.. Differential association among Timp1, CD63 and β1-integrins was observed in melan-a melanocytes, 4C pre-malignant melanocytes and 4C11- and 4C11+ melanoma cells. Timp1 present in conditioned medium of melanoma cells rendered melan-a melanocytes anoikis-resistant through PI3-K signaling pathway independently of Akt activation. In human melanoma cell lines, in which TIMP1 and beta-1 integrin were also found to be interacting, TIMP1 and CD63 levels together was shown to correlate significantly with colony formation capacity.. Our results show that Timp1 is assembled in a supramolecular complex containing CD63 and β1-integrins along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway, independently of Akt phosphorylation. In addition, our data point TIMP1, mainly together with CD63, as a potential biomarker of melanoma.

Topics: Anoikis; Cell Line; Cell Survival; Cell Transformation, Neoplastic; Culture Media, Conditioned; Disease Progression; Gene Expression; Humans; Integrin beta1; MART-1 Antigen; Melanocytes; Melanoma; Models, Biological; Multiprotein Complexes; Neoplasm Metastasis; Phenotype; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Protein Binding; Proto-Oncogene Proteins c-akt; Signal Transduction; Tetraspanin 30; Tissue Inhibitor of Metalloproteinase-1

Intraocular melanocytoma is a rare naevus variant that can be located at the optic disc or within the uvea, and belongs to the group of non-epithelial-associated melanocytic lesions. We wanted to gain an understanding of the role of GNAQ, GNA11 and BRAF V600E in the pathogenesis of uveal melanocytoma and in cases of transformation to uveal melanoma and also to perform a differential immunohistochemical study comparing melanocytoma with uveal melanoma.. Two patients were identified with melanocytoma, one of which had transformed to melanoma. In the latter case, the melanocytoma exhibited an immunophenotype that featured nuclear p27 and no HMB45 staining, with very low Cyclin D1 expression compared with the melanoma that featured little nuclear but more cytoplasmic p27 positivity, much higher Cyclin D1 expression and HMB45 positivity. The melanocytomas were negative for CD68 allowing distinction from melanophages. Both melanocytomas and the melanoma harboured mutations in GNAQ, with no mutations of GNA11 or BRAF V600E.. GNAQ mutations are present in uveal melanocytomas and in a case of transformation to melanoma, implicating GNAQ-dependent mitogen activation signals, in the pathogenesis of uveal melanocytoma. This assists in explaining why a proportion of uveal melanocytoma can transform to uveal melanoma, known to harbour high-frequency GNAQ mutations at exon 5, codon 209.

Topics: Adolescent; Adult; Cell Transformation, Neoplastic; Chromosomes, Human, Pair 3; Chromosomes, Human, Pair 8; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p27; gp100 Melanoma Antigen; GTP-Binding Protein alpha Subunits; GTP-Binding Protein alpha Subunits, Gq-G11; Humans; Immunoenzyme Techniques; In Situ Hybridization, Fluorescence; Male; MART-1 Antigen; Melanoma; Melanoma-Specific Antigens; Mutation; Nevus, Pigmented; Polymerase Chain Reaction; Proto-Oncogene Proteins B-raf; Retrospective Studies; Uveal Neoplasms

Topics: Adolescent; Biomarkers, Tumor; Biopsy; Cell Transformation, Neoplastic; Conjunctival Neoplasms; gp100 Melanoma Antigen; Humans; Male; MART-1 Antigen; Melanoma; Melanoma-Specific Antigens; Nevus, Pigmented; S100 Proteins

This study explored the possibility that the components in melanoma cytoplasm induce murine BMSCs transformation and expression of Melan-A by morphologically observing the changes of BMSCs and immunocytochemically detecting Melan-A in the cells after culturing BMSCs in medium containing melanoma cytoplasm components (MCC). MCC of B16 melanoma cells was prepared and BMSCs were cultured and induced by adding the MCC into culture medium. The cells were morphologically observed and Melan-A was immunohistochemically detected to confirm BMSCs transformation. MCC-induced BMSCs underwent morphological changes. A number of melanin granules appeared in the cytoplasm of the cells and some were released into surrounding areas. Several cells that might come from one cell formed a cluster, and their granules, together with those secreted by other induced BMSCs, formed a so-called "sphere-formed structure". The induced BMSCs expressed Melan-A. We are led to conclude that there might be some factors in the cytoplasm of melanoma cells that might induce BMSCs transformation toward melanogenic cell, or even melanoma.

Topics: Animals; Bone Marrow Cells; Cell Line, Tumor; Cell Transformation, Neoplastic; Coculture Techniques; Cytoplasm; Humans; Male; MART-1 Antigen; Melanoma, Experimental; Mice; Skin Neoplasms; Stem Cells

Topics: Adenoma; Adrenal Cortex Neoplasms; Aged; Antigens, Neoplasm; Biomarkers, Tumor; CD56 Antigen; Cell Dedifferentiation; Cell Transformation, Neoplastic; Female; Humans; MART-1 Antigen; Neoplasm Proteins; Synaptophysin

Topics: Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinosarcoma; Cell Transformation, Neoplastic; Female; Humans; Lymph Nodes; MART-1 Antigen; Melanoma; Melanoma-Specific Antigens; Neoplasm Proteins; S100 Proteins; Skin Neoplasms

To report the protocol for surveillance of the eye and vision in a clinical trial of MART1-transduced dendritic cells for metastatic melanoma.. In a phase I/II clinical trial of dendritic cell-based genetic immunotherapy for metastatic cutaneous melanoma, ophthalmic evaluation is performed prior to immunization (Baseline Evaluation), 56+/-7 days after first vaccination (mid-study evaluation), when dendritic cell injections are complete 112+/-7 days after first vaccination (end-study evaluation) and 168+/-7 days after first vaccination (post-study evaluation).. The protocol for baseline, mid-study and end-study evaluations of the eye and vision includes ophthalmic history, comprehensive ophthalmic examination, psychophysical and electrophysiological visual function assessment, fundus photography and fluorescein angiography. Post-study evaluation consists of the 25-item visual functioning questionnaire augmented to elicit autoimmune manifestation with complete ophthalmic evaluation if vision-related symptoms or abnormalities are noted during or after the vaccination.. Limited adverse effects on the eye and vision have been reported in melanoma immunotherapy trials, although this novel mode of therapy has the potential to induce melanoma paraneoplastic syndromes known to severely impair vision. Therefore, surveillance of the eye and vision should be considered in melanoma immunotherapy trials.

Topics: Antigens, Neoplasm; Cancer Vaccines; Cell Transformation, Neoplastic; Clinical Trials as Topic; Contrast Sensitivity; Dendritic Cells; Dose-Response Relationship, Immunologic; Electroretinography; Eye Diseases; Genetic Therapy; Humans; Immunotherapy; Injections, Intradermal; MART-1 Antigen; Melanoma; Neoplasm Proteins; Population Surveillance; Skin Neoplasms; Transduction, Genetic; Vaccination; Vision Disorders; Visual Acuity; Visual Fields

A panel of three melanocyte differentiation antibodies has been compared with anti-S100 protein and NKIC3 in an assessment of benign and malignant melanocytic lesions. Anti-polyclonal S100 protein labelled all cases of primary cutaneous malignant melanoma, metastatic melanoma, desmoplastic melanoma and myxoid melanomas. In addition all benign and dysplastic naevi were positive. Conversely, HMB 45 was the least sensitive marker, labelling 24/31 primary cutaneous melanomas, 14/24 metastatic melanomas and only 1/6 desmoplastic melanomas. In the case of naevi, only junctional forms labelled consistently. Results for anti-melan-A and anti-tyrosinase were similar, although anti-tyrosinase proved slightly more sensitive in cases of malignant melanoma. NKIC3 revealed similar results to anti-tyrosinase, but had the disadvantage of reduced selectivity. It is concluded that anti-tyrosinase and anti-melan-A are useful additions to the panel of melanocytic monoclonal antibodies. In addition, both antibodies appear to have greater sensitivity for malignant melanoma than the conventionally used HMB 45 and could be considered as supportive markers to polyclonal anti-S100 protein in the diagnosis of malignant melanoma.

Topics: Antibodies, Monoclonal; Antibody Specificity; Antigens, Neoplasm; Biomarkers, Tumor; Cell Differentiation; Cell Transformation, Neoplastic; Humans; Immunohistochemistry; MART-1 Antigen; Melanocytes; Melanoma; Melanoma-Specific Antigens; Monophenol Monooxygenase; Neoplasm Proteins; S100 Proteins