Expression analysis of the transcripts and proteins COX-2, AKT, NFKB, IKB, MTOR and CD1 in cell cultures of Head and Neck squamous cell carcinomas treated by CELECOXIB

• Grant number: 07/00870-4
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2007
• End date: March 31, 2010
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Decio dos Santos Pinto Junior
• Grantee: Decio dos Santos Pinto Junior
• Host Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Several genetic alterations are related to squamous cell carcinoma (SCC) pathogenesis, the most frequent malignancy in head and neck. Some of these alterations are involved in AKT signaling pathway associated in several cellular events. COX 2 protein initially described in inflammation is also related with AKT pathway. This study aims to verify the pAKT, NF-kB, IKB, mTOR, ciclyn D1 e COX-2 proteins and transcripts in squamous cell carcinoma cell lines to verify alterations in transduction and translation under EGF stimulation and CELECOXIB treatment. Four squamous cell carcinoma lines will be divided in three different groups: a- control, b- treated with Celecoxib and c- stimulated with EGF and treated with Celecoxib. The cellular proliferation and viability will be monitored with growth curves and cell viability assay. Protein analysis will be realized by Western Blotting technique and proteins interactions by co-immunoprecipitation. Immunofluorescence technique will be used to verify the intracellular protein location and the apoptotic cells by TUNEL. Transcripts quantification will be performed by Real Time PCR. By these techniques the objective of this study is to have better visualization concerning the Celecoxib action in AKT pathway in SCC. (AU)