Role of microRNA-9 in control of Nox4 expression and in migration of melanoma cells

• Grant number: 17/24439-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): June 01, 2018
• Effective date (End): December 31, 2018
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Lucia Rossetti Lopes
• Grantee: Marcella Liciani Franco
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 13/07937-8 - Redoxome - Redox Processes in Biomedicine, AP.CEPID

Abstract

NADPH oxidase 4 (Nox4) is an isoform of NADPH oxidase. This family of enzymes is dedicated to the production of reactive oxygen species (ROS) in cells. It is known that, in melanoma cells, the expression and activity of Nox4 are increased when compared to healthy cells and it may influence the proliferation, migration and survival of malign cells. Since melanoma is the most aggressive type of skin cancer, especially due to its metastatic capacity, elucidating cellular pathways which regulate Nox4 expression in this tumour is relevant so that new therapeutic targets can be found, for instance. Studies suggest that a possible regulation pathway of this enzyme is through miR-9, a microRNA with altered expression in melanoma cells. Therefore, the aim of this project is to investigate whether miR-9 modifies the migration of melanoma cells through the regulation of Nox4 expression. We will use some techniques such as culture of melanocytes (NGM lineage) and of melanoma cells (A-375, SK-Mel 19, SK-Mel28 and SK-Mel 147 lineages), extraction of their total RNA and RT-qPCR for Nox4 and miR-9. In addition, we will perform both Western blot essays to evaluate the expression of Nox4 protein in these cells and transfection of them with a miR-9-mimic so that it will be overexpressed. After transfection, new quantifications of Nox4 expression and migration essays will be fulfilled. (AU)