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Communication Dans Un Congrès Année : 2011

High-throughput mRNA profiling of microdissected melanocytes and melanoma cells for the characterization of melanoma predisposition in a swine model

Résumé

Background: Melanoblastoma-bearing Libechov Minipigs (MeLiM) present spontaneous tumors at birth or at early ages that share many clinical and histopathological features with human cutaneous melanoma (1). Remarkably, in swine, as in humans and contrary to mice, the melanocytes are located in the basal cell layer of the epidermis. Thus, MeLiM pigs stand as an exceptional model for human hereditary melanoma and we use them to study the genes involved in oncogenesis. Objectives: To our knowledge, in humans, all gene expression studies on melanocytes have been performed by using cultured cells, in which the transcript levels could be dramatically modified by the culture conditions. To overcome this limitation, the objectives of this study were to develop and optimize a protocol for single-cell laser capture microdissection (LCM) of skin melanocytes from tissue sections by using an Arcturus Veritas LCM system. Methods: To identify genes that may regulate the malignant transformation of melanocytes, the expression profiles of swine microdissected melanocytes and melanoma cells were compared between healthy and affected MeLiM-derived pigs by using high-throughput RNA sequencing (RNA-seq). Results-Discussion: Given that the amplification step in the LCM protocol creates a 3 prime bias, we have adapted current bioinformatic analyses to better estimate the expression levels of genes. Our protocol uses a window of 250 bases from the 3 prime end of the transcript. Additionally, expressed single nucleotide variants have been identified. The results obtained will help to understand the melanoma predisposition not only in MeLiM pigs but also in humans.

Domaines

Cancer
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Dates et versions

hal-01019847 , version 1 (07-07-2014)

Identifiants

  • HAL Id : hal-01019847 , version 1
  • PRODINRA : 175094

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Jordi Estellé, Guillaume Piton, Claudia Bevilacqua, Stephan Bouet, Emmanuelle Bourneuf, et al.. High-throughput mRNA profiling of microdissected melanocytes and melanoma cells for the characterization of melanoma predisposition in a swine model. 7th EADO Congress, Jun 2011, Nantes, France. ⟨hal-01019847⟩
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