NOVA1 RNAi silencing in glioblastoma cell lines: proteomic analysis of the targets of this splicing factor and its repercussions on gliomagenesis

• Grant number: 17/09542-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2017
• End date: December 31, 2018
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: José César Rosa
• Grantee: Igor Fernando Almeida Teodoro
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The alternative splicing, in the context of the diversity of protein products from a restricted number of genes, carries great importance for an understanding of the mechanisms that govern the cellular physiology of different tissues. In this sense an understanding of how mechanism is regulated, especially by the sayings of splicing, becomes important. Among them, as proteins of the NOVA family, mainly NOVA1, considering as discoveries relating this factor of regulation of various cell types, such as beta-pancreatic cells and neural cells. As with other splicing factors, a change in NOVA1 expression has been related to the development of neoplasms. Our group has recently shown to have a large difference in factor expression between neoplastic and non-neoplastic neural tissue cells, justifying an investigation of NOVA1 function. To achieve this goal, NOVA1 will be silenced by siRNA and the effect of this generic silencing will be investigated by proteomic techniques in glioblastoma strains. 1) RNAi silencing _NOVA1; 2) Cell proliferation assay and cell viability; 3) mRNA of NOVA1 expression by RT-PCR and abundance of NOVA1 by Western Blotting and 4) proteomic analysis of the silenced cells versus parental and NTC cells. (AU)