Study of the role of OR2C3 in melanoma development and progression

  • Clémence François

Student thesis: Master typesMaster en sciences biomédicales à finalité spécialisée en recherche préclinique

Résumé

Background – Apart from their main expression in olfactory sensory neurons, olfactory receptors (ORs) have also been found expressed in a wide range of other tissues, suggesting the existence of other biological functions. Interestingly, ORs were shown to be involved in some cancerous processes. A well-known example is the implication of OR51E2 in prostate cancer. Recently, Ranzani and colleagues led a study about the expression of ORs in various cancer cell lines, which revealed OR2C3 as the only OR expressed in melanoma, while no expression was detected in normal melanocytes. These data suggest that OR2C3 may contribute to the development and the progression of melanoma.
Aim – The aim of this master thesis was to generate preliminary data over the potential role of OR2C3 in melanoma development and progression.
Methods – In silico studies were carried out using various software. RT-qPCR was used to evaluate the level of OR2C3 mRNA expression. Then selected cell lines were transfected with a Rho-OR2C3 plasmid, and the mRNA and protein expression levels of Rho-OR2C3 were assessed by RT-qPCR and western blot. Immunofluorescence was also performed to determine the localization of the receptor. Finally, functional assays were conducted including a proliferation assay and a soft agar colony formation assay.
Results – Bioinformatics allowed to establish a clearer representation of OR2C3 at the genetic and the protein levels. RT-qPCR revealed that MEL-JU and MEL-JUSO melanoma cell lines express OR2C3 at the greatest level among all tested melanoma cell lines. Since no antibody is available to detect the native protein, transfection of Rho-tagged OR2C3 was successfully performed in MEL-JU and MEL-JUSO. Rho-tagged OR2C3 was detectable at the mRNA and protein levels. Immunofluorescence revealed that Rho-tagged OR2C3 was mainly found in the Golgi apparatus. Proliferation and colony formation assays did not show any significant difference between studied conditions.
Conclusion – The overexpression of Rho-OR2C3 in MEL-JU and MEL-JUSO cell lines did not reveal any changes in cells behavior. However, immunofluorescence also revealed that Rho-tagged OR2C3 localization was not found predominantly at cell membrane, as we might expect. This indicate the need to transfect the RTP1S accessory protein to facilitate the OR2C3 trafficking to the cell-surface membrane.
la date de réponse17 janv. 2022
langue originaleAnglais
L'institution diplômante
  • Universite de Namur
SuperviseurJean-Pierre Gillet (Promoteur)

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