Evaluation of the role of miRNAs in vulvar carcinomas: from tumor expression profile to in vitro studies

• Grant number: 12/13832-1
• Support Opportunities: Regular Research Grants
• Start date: March 01, 2013
• End date: February 28, 2015
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Rafael Malagoli Rocha
• Grantee: Rafael Malagoli Rocha
• Host Institution: A C Camargo Cancer Center. Fundação Antonio Prudente (FAP). São Paulo , SP, Brazil

Abstract

Vulvar carcinoma is a rare gynecologic tumor whose lack of biological information justifies the necessity of novel biomarker Discovery. These new findings allow the comprehension of the important mechanisms in this type of tumor, such as miRNAs, whose disregulated expression has been proved to be important in several tumorigenesis processes. Aims: Characterize miRNA expression profile in vulvar cancer samples in the presence or not of high risk HPV infection. Methods: Fourty samples of squamous cell carcinoma of the vulva had been already selected, among those, 20 samples are HPV positive and 20 are HPV negative. Ten normal samples of tumor border will be microdissected in order to generate a pool of global non-tumoral control. The identification of the differentially expressed miRNA will be evaluated by RT-PCR using the card system TaqMan human microRNA array 2.0 A+B. It will also be performed miRNA validation in frozen tissue, in silico search of possible target genes (using algorithms such as miRWalk, miRanda, PicTar, miRBase and TargetScan), biological activity tests of the selected miRNAs by using vulvar tumor cell culture (SW-954), and also evaluate gene expression of those genes which are regulated by the selected miRNAs of this study. It will also be performed in large-scale gene expression, immunohistochemistry, immunofluorescence and immunocitofluorescence in order to compare molecular, morphological and clinical results. (AU)