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Evaluation of the diagnostic potential of genes belonging to the MAP/KINASE pathway in thyroid lesions

Grant number: 19/10211-5
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): August 01, 2019
Effective date (End): July 31, 2020
Field of knowledge:Biological Sciences - Genetics - Human and Medical Genetics
Principal Investigator:Renata de Azevedo Canevari
Grantee:João Paulo da Silva Queiroz
Home Institution: Instituto de Pesquisa e Desenvolvimento (IP&D). Universidade do Vale do Paraíba (UNIVAP). São José dos Campos , SP, Brazil

Abstract

Thyroid cancer is the most common pathology affecting the endocrine system, and there has been an increase in the number of cases diagnosed in recent years. The MAP/KINASE signaling pathway has been well described as a diagnostic target for lesions affecting the gland due to genetic changes related to the development and progression of tumors. Diagnosis is obtained by the association of two methods: ultrasonography and fine needle aspiration. However, due to the low sensitivity, both have a high rate of inconclusive results, making it necessary to search assiduously for alternative diagnostic methods and with more accurate results. Molecular biology has been prominent in the search for genetic mutations and alterations of gene expression to enable the identification of potential diagnostic markers in the different lesions. Thus, this study aims to analyze the expression of the CDKN1C, CREB1, FOS, HSPA5, JUN, KSR1, MAP2K6, MAPK8IP2 and SFN genes belonging to the MAP/KINASE pathway as potential diagnostic markers. These genes will be evaluated in 50 samples of thyroid tissues: 10 samples of PTC, 10 of FTC, 10 of adenomatous goiter, 10 of benign tumors (5 of follicular adenoma and 5 of follicular hyperplasia) and 10 of non-tumor tissue ( No. 1,806,781 / CEP / 2016). The RNeasy® Mini kit and SuperScript² IV First-Strand Synthesis System protocols will be used for RNA extraction and cDNA synthesis, respectively. Gene expression analysis by the quantitative real-time PCR technique (RT-qPCR) will be performed using the SYBR Green qPCR Master Mix RT² on the ABI Prism 7500 Sequence Detection Systems. The data obtained will be translated by the software Sequence Detection System software 2.1. Statistical analyzes will be performed on GraphPad Prism 5.00 software, including the non-parametric Mann Whitney test (p d 0.05). As a result, it is expected to identify potential molecular markers of the MAP/KINASE pathway in order to assist in a more accurate diagnosis of these lesions.