Evaluation of the role of microRNA in vulvar carcinoma

Abstract

Background: Vulvar carcinoma is a rare gynecologic tumor in which the lack of biological information becomes strongly the need to identify molecules to enable the understanding of the mechanisms of this type of tumor, such as microRNAs, which dysregulated expression has been demonstrated as one of the major determinants of tumorigenesis. Objectives: To characterize the expression profile of miRNAs in samples of vulvar carcinomas in the presence and absence of high-risk HPV. Methods: Forty samples of squamous cell carcinoma of the vulva will be selected, with HPV 20 positive samples and 20 HPV negative. Ten samples from microdissected normal tissue will be used to build a pool to be used as a non-tumor overall control. The identification of differentially expressed miRNAs in the different samples will be performed by qRT-PCR using the card system TaqMan Array Human MicroRNA 2.0 A + B. There will be also carried out the validation of selected miRNAs in fresh tissue, the in silico search of potential target gene of these miRNAs by using algorithms such as miRWalk, Miranda, PicTar, miRBase and TargetScan, and testing of biological activity of miRNAs elected in a vulvar cell culture (SW-954), as well as the evaluation of the gene expression of targets of these miRNAs by qRT-PCR and protein expression by immunocytochemistry.