Activation of the PI3K pathway in the expression of FBLN1 and CLDN6 by triiodothyronine and estradiol in a breast carcinoma cell line

• Grant number: 14/23786-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2015
• End date: November 30, 2016
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Sandro José Conde
• Grantee: Gabriela Silva de Souza
• Host Institution: Instituto Federal de Educação, Ciência e Tecnologia de São Paulo (IFSP). Campus São Roque. São Roque , SP, Brazil

Abstract

Recently our group demonstrated FBLN1 and the CLDN6 genes have been stimulated by E2 and T3 in MCF-7 cells. Thus,our hypothesis is that thyroid hormone alters the expression of FBLN1 and CLDN6 without the need of translocation to the nucleus, by activation of phosphatidylinositol-3-kinase (PI3K) pathway. We intend to elucidate the pathway of action of E2 and T3 in the activation of these gene targets in cell lines of breast adenocarcinoma. It will be used inhibitor LY294002 (50¼M), which blocks the PI3K pathway and shows us if this action is dependent on this pathway. Methods: Cell lines of breast cancer MCF-7 will plated at intervals of 10 minutes, 30 minutes, 1 hour and 4 hours E2 (10-7M), T3 (10-8M), E2 (10-7M) + LY294002 (50¼M), T3 (10-8M) + LYLY294002 (50¼M) and LY294002 (50¼M). All treatments will be performed in triplicate. It will be performed total RNA extraction to obtain cDNA - Reverse transcription (RT) of RNA and Real time PCR for expression of genes in each treatment.