Haplosufficiency of PAX3 for melanoma development in Tyr: NRASQ61K; Cdkn2a-/- mice allows identification and sorting of melanoma cells using a Pax3GFP reporter allele

Autor: Jacky Ezagal, Geneviève Aubin-Houzelstein, Friedrich Beermann, Cécile Campagne, Giorgia Egidy, Stéphanie Gadin, Jean-Jacques Panthier, Marie Abitbol, Florence Bernex, Sophia Loiodice, Edouard Reyes-Gomez, Anne Louise
Přispěvatelé: Génétique fonctionnelle et médicale, École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Unité d'Embryologie, Histologie et Anatomie Pathologique, École nationale vétérinaire - Alfort (ENVA), Institut de Recherche en Cancérologie de Montpellier (IRCM - U1194 Inserm - UM), CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), BioCampus (BCM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Cytométrie (Plate-forme), Institut Pasteur [Paris] (IP), Swiss Institute for Experimental Cancer Research, ISREC, Génétique fonctionnelle de la Souris, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), This work was supported by grants from the Institut National de la Recherche Agronomique, the Agence Nationale de la Recherche Emergence Bio and the Association pour la Recherche contre le Cancer. CC was granted by Allocation de Recherche MENRT from the French Ministry of Research, ANR-09-EBIO-0006,blackRACK1,Evaluation de l'intérêt diagnostic de RACK1 dans les mélanomes(2009), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), École nationale vétérinaire d'Alfort (ENVA), BioCampus Montpellier (BCM), Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Rok vydání: 2015
Předmět:
0301 basic medicine
Neuroblastoma RAS viral oncogene homolog
Cancer Research
Pathology
Skin Neoplasms
[SDV]Life Sciences [q-bio]
Cell Separation
Haploinsufficiency
Green fluorescent protein
Mice
0302 clinical medicine
CDKN2A
Genes
Reporter
Paired Box Transcription Factors
Melanoma
Cells
Cultured
Mice
Knockout
primary culture
Monophenol Monooxygenase
Cell sorting
musculoskeletal system
3. Good health
Cell Transformation
Neoplastic
Oncology
direct FACS-sorting
030220 oncology & carcinogenesis
embryonic structures
Immunohistochemistry
Female
medicine.medical_specialty
mouse melanoma models
Green Fluorescent Proteins
Dermatology
Biology
03 medical and health sciences
In vivo
medicine
Animals
neoplasms
PAX3 Transcription Factor
Alleles
PAX3
Genes
p16
immunohistology
medicine.disease
Embryonic stem cell
Mice
Inbred C57BL
030104 developmental biology
Genes
ras
Amino Acid Substitution
Cancer research
Zdroj: Melanoma Research
Melanoma Research, 2016, 26 (1), pp.12-20. ⟨10.1097/CMR.0000000000000212⟩
Melanoma Research, Lippincott, Williams & Wilkins, 2016, 26 (1), pp.12-20. ⟨10.1097/CMR.0000000000000212⟩
ISSN: 1473-5636
0960-8931
DOI: 10.1097/CMR.0000000000000212⟩
Popis: International audience; The role of the Pax3 gene in embryonic development of pigment cells is well characterized. By contrast, the function of Pax3 in melanoma development is controversial. Indeed, data obtained from cultured cells suggest that PAX3 may contribute to melanomagenesis. PAX3 is found to be overexpressed in melanomas and also in nevi compared with normal skin samples. Pax3 homozygous loss of function is embryonic lethal. To assess the role of Pax3 in melanoma development in vivo, we analyzed Pax3 haploinsufficiency in a mouse model of melanoma predisposition. The Pax3(GFP/+) knock-in reporter system was combined with the Tyr::NRAS(Q61K); Cdkn2a(-/-) mouse melanoma model. Melanoma development was followed over 18 months. Histopathological, immunohistochemical, and molecular analyses of lesions at different stages of melanoma progression were carried out. Fluorescence-activated cell sorting on GFP of cells from primary or metastatic melanoma was followed by ex-vivo transformation tests and in-vivo passaging. We report here that Tyr::NRAS(Q61K); Cdkn2a(-/-); Pax3(GFP/+) mice developed metastasizing melanoma as their Tyr::NRAS(Q61K); Cdkn2a(-/-); littermates. Histopathology showed no differences between the two genotypes, although Pax3 mRNA and PAX3 protein levels in Pax3(GFP/+) lesions were reduced by half. The Pax3(GFP) allele proved to be a convenient marker to identify and directly sort heterogeneous populations of melanoma cells within the tumor bulk at each stage of melanoma progression. This new mouse model represents an accurate and reproducible means for identifying melanoma cells in vivo to study the mechanisms of melanoma development.
Databáze: OpenAIRE
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