Characterization of Gja1 gene expression using a response dose curve in pulmonary neoplasic cells transfected with induclible system drived by doxiciclin

• Grant number: 14/11376-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2014
• End date: July 31, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Pathology
• Principal Investigator: Lucas Martins Chaible
• Grantee: Ana Paula Dias
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Lung cancer is a major cause of death in modern society , responsible for over 10 million deaths per year worldwide, and its incidence is directly related to the habits of the day -to-day , between him the habit of smoking .The cell line E9 corresponds to the epithelial tumor cells from murine lung. These cells naturally exhibit decreased expression of Gja1 gene responsible for the protein Cx43 intercellular communication. This reduction in intercellular communication has been correlated with higher malignancy of tumors because tumor clones are not removed by the microenvironment surrounding them. The purpose of this job is to be a step further in another project of gene therapy using gene Gja1. In this major work, we aim to see if the increased expression of Cx43 gene causes the restoration of intercellular communication and consequently the change in the characteristics of malignancy of cancer cells . In this research project we aim to standardize the dose response curve between doxycycline and referring expression level Gja1 gene, which is present in the conditional expression vector for the drug. To achieve this objective, lung tumor cells E9 , where by transfection of inducible gene expression by doxycycline drug ( Teton - Cx43 ) system will stimulate the expression of the heterologous protein . This expression system is already built and tested, and it was observed that the presence of this antibiotic the system is activated and Gja1 gene is expressed, its expression is turned off with the withdrawal of the drug. This project aims precisely to relate doses of doxycycline (100ng/ml, and 500ng/mL 1000ng/ml) with the expression levels of the gene Gja1 quantified by Real Time PCR technique.