Expression and Functional Characterization of the BNIP3 Protein in Renal Cell Carcinomas

Background. Bilitranslocase (TC 2.A.65.1.1) is a bilirubin-specific membrane transporter, found on absorptive (stomach and intestine) and excretory (kidney and liver) epithelia and in vascular endothelium. Polyclonal antibodies have been raised in rabbits in the past, using a synthetic peptide corresponding to AA65-77 of rat liver bilitranslocase, as an antigen. Affinity-purified antibodies from immune sera have been found to inhibit various membrane transport functions, including the bilirubin uptake into human hepatocytes and the uptake of some flavonoids into human vascular endothelial cells. It was described by means of immunohistochemistry using polyclonal antibodies that bilitranslocase expression is severely down-regulated in clear cell renal carcinoma. The aim of our work was development and characterization of high-affinity, specific mAbs against bilitranslocase, which can be used as a potential diagnostic tool in renal cell carcinoma as well as in a wide variety of biological assays on different human tissues. Materials and methods. Mice were immunized with a multi-antigen peptide corresponding to segment 65-75 of predicted primary structure of the bilitranslocase protein. By a sequence of cloning, immune- and functional tests, we aimed at obtaining a specific monoclonal antibody which recognizes a 37 kDa membrane protein, and influences the transport activity of bilitranslocase. Results. On the basis of previous results, specific IgM monoclonal antibodies were produced in BALB/c mice, in order to further improve and extend the immunological approach to the study of bilitranslocase in renal cancer cells as well as to develop its potential diagnostics use. Conclusions. In this article we show an immunological approach, based on newly developed monoclonal antibodies, to a detailed biochemical and functional characterization of a protein whose gene and protein structure is still unknown. We were able to demonstrate our novel mAb as a tumor marker candidate of renal cell carcinoma, which may prove useful in the diagnostic procedures.

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Characterization of quantitative chromosomal abnormalities in renal cell carcinomas by interphase four-color fluorescence in situ hybridization

Cancer Genetics and Cytogenetics ◽

10.1016/j.cancergencyto.2004.08.019 ◽

2005 ◽

Vol 158 (2) ◽

pp. 110-118 ◽

Cited By ~ 18

Author(s):

Aline Ossard Receveur ◽

Jérôme Couturier ◽

Vincent Molinié ◽

Annick Vieillefond ◽

François Desangles ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Renal Cell ◽

Chromosomal Abnormalities ◽

Renal Cell Carcinomas

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VHL-mutant renal cell carcinomas contain cancer cells with mesenchymal phenotypes.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.4568 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. 4568-4568 ◽

Cited By ~ 2

Author(s):

Craig Gedye ◽

Danylo Sirskyj ◽

Nazleen Carol Lobo ◽

Andrew Evans ◽

Neil Eric Fleshner ◽

...

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

Cancer Cells ◽

Tumor Cells ◽

Renal Cell ◽

Clear Cell ◽

Ex Vivo ◽

Functional Characterization ◽

Mutant Cells

4568 Background: To study “cancer stem cells” it is imperative to account for all stromal cell populations within the tumour. The existence of “cancer stem cells” in clear cell renal cell carcinoma (ccRCC) has not been examined in ex vivo patient samples. Methods: We established a multiplex flow cytometry (FC) antibody panel in ccRCC, which reliably identified stromal lineages including CD45+ immune, CD31+/CD144+ endothelial and fibroblast-marker-positive subpopulations, thus allowing isolation of "lineage-negative" tumor cells. To verify the identity of tumour-derived populations as either cancer cells or normal stromal cells, we took advantage of the fact that mutations in VHL occur early during ccRCC tumorigenesis and are found in two-thirds of patients. Results: We sequenced 18 patient tumor samples, 12 of which had VHL exome mutations. Targeted re-sequencing of FC sorted subpopulations from these patients’ samples revealed that while CD45+ immune cells and CD31+/CD144+ endothelial cells were genetically normal, a population of VHL-mutant fibroblast-marker positive cells was consistently identified in every patient’s tumour. Immunohistochemistry showed that fibroblast marker-positive VHL-mutant cells do not have the large “clear cell” morphology typical of the majority of the cancer cells in these tumours. When purified and cultured, these fibroblast marker-positive VHL-mutant cells proliferate extensively under mesenchymal culture conditions, but displayed different morphologies to lineage-negative VHL-mutant tumor cells. Functional characterization of these FC sorted cell subpopulations is ongoing, including proliferation, migration, invasion, differentiation and treatment resistance. Conclusions: The phenotype and preliminary functional characterization of these VHL-mutant fibroblast-marker positive cells suggests a mesenchymal differentiation program in ccRCC, with implications for the ontogeny, biology and clinical management of VHL-mutant renal cancer.

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9º Simpósio Médico Internacional de VHL; III Encontro de Famílias com a Síndrome de VHL Data:

Revista Brasileira de Cancerologia ◽

10.32635/2176-9745.rbc.2010v56n3.1493 ◽

2010 ◽

Vol 56 (3) ◽

pp. 395-406

Author(s):

Instituto Nacional de Câncer José de Alencar Gomes da Silva

Keyword(s):

Renal Cell Carcinoma ◽

Tumor Suppressor ◽

Cell Carcinoma ◽

Renal Cell ◽

Renal Cell Carcinomas ◽

Vhl Gene ◽

Von Hippel Lindau ◽

Number Variation ◽

Von Hippel Lindau Disease

Resumos escolhidos para publicação. Nessa edição, os títulos foram: Extramitochondrial Fumarate Inhibits Multiple 2-OG Oxygenases in Fumarate Hydratase Deficient Cells; What is the Best Treatment for Renal Lesions in VHL?; Characterization of the VHL-ECM Pathway; Induction of Extreme Metabolic Depression by the Nucleolus; Mutation of SDHB and Inherited RCC Susceptibility; Metabolic Links to Renal Cancer; Radiosurgery for Cerebellar Hemangioblastomas; Neoplastic Diagnosis Timing Profile in von Hippel-Lindau´s Patients in a Personal Series; Regulation of the VHL Tumor Suppressor; Destructive Targeting via VHL Beyond HIF; Folliculin Functional Studies and Mouse Models of Birt-Hogg-Dube’ Syndrome; A Zebrafish Model for VHL; Relief of Intractable Nausea after Resection of Brainstem Hemangioblastoma in Patients with von Hippel-Lindau Disease: a Clinical Series; Somatic Alteration of the VHL Gene in Sporadic Renal-Cell Carcinomas as a Potential Biomarker; VHL Tumor Suppressor Protein Regulates Oncogenic Macroautophagy in Renal Clear Cell Carcinoma (RCC); Identification of Germline Mutations in the VHL Gene of Families with the von Hippel-Lindau Disease; Expression Profile in von Hippel-Lindau Disease Associated and Sporadic Clear-Cell Renal Cell Carcinomas; Proposed Changes to the VHL Handbook; Evaluation of the Somatic Alterations of the VHL Gene in Renal Cell Carcinoma Associated with von Hippel-Lindau Disease (VHL); Copy Number Variation Analysis of a Pancreatic Neuroendocrine Tumor (NET) from a Patient with von Hippel-Lindau (VHL); Molecular Dynamics Study of the Mutant pVHL Phe76del and its Interactions with Components of the pVHL Complex; Genomic Copy Number Variation Analysis in VHL-Associated Renal Cell Carcinomas Suggests Clonality; Management of Brainstem and Spinal cord Hemangioblastomas in Patients with von Hippel-Lindau Disease; The Benefits of Ultrasound in Resection of CNS Hemangioblastomas; Management of Central Nervous System Hemangioblastomas in von Hippel-Lindau Disease; Neuronal Differentiation of Stem cells by Transfer of a VHL Peptide and Regenerative Therapy; Endolymphatic Sac Tumors (ELST) in VHL Patients - Evaluation of Screening Methods in a National Study; Delineating Genotype-Phenotype Correlations Among Brazilian Families with von Hippel-Lindau Disease; Endothelial Fenestrations Associated with VHL Gene Alteration is a Potent Target of Anti-VEGF Therap; Role of Pregnancy on Hemangioblastomas in von Hippel-Lindau Disease: a Retrospective French Study; An Evaluation of the Danish National Clinical Guidelines for Von Hippel-Lindau (VHL); Vitreoretinal Surgery for Severe Retinal Capillary Hemangiomas; Oncololytic Targeting of Renal Cell Carcinoma via Encephalomyocarditis Virus; Emerging Therapeutic Options for VHL Patients: a Tale of three Studies; Altering the Stability of Mutant VHL: Potential Therapeutic Consequences; Tale of the Tail: Clinical and Functional Properties of Novel VHL Mutation (X214L) Consistent with Type 2A Phenotype and Low Risk of Renal Cell Carcinoma; Knife for Intracranial Hemangioblastomas in von Hippel-Lindau Patients. When and How?; VHL in Brazil: Genetics, Biobanking and VHL Family Care; Understanding VHL Disease: Molecular Characterization of a Spanish Series; Case Report: Radiosurgery for Endolymphatic Sac Tumor in a Patient with von Hippel-Lindau Disease; Primary Cilium: a Tumor Suppressor Organelle.

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