Differentially expressed gene assessment in vulvar cancer: validations based in results of comparative genomic hybridization array

Abstract

Vulvar carcinoma is a rare disease that accounts for less than 3% of all female cancers. In younger women this cancer is closely linked to smoking and HPV infection but the prevalence of vulvar cancer cases occur in women over 50 years by genetic mechanisms that still poorly understood. In the past few years some studies showed genomic alterations in vulvar carcinomas, however none of them determinate any new prognostic factor or correlated these findings with gene expression profiles. From the analysis of CGH-array conducted by PhD Student André Mourão Lavorato da Rocha in our laboratory, two candidate genes, LIMK1 and ROCK1, highly expressed in vulvar carcinomas were selected to evaluate its importance in this type of cancer. Aims: To evaluate, by Immunohistochemistry and qRT-PCR, the expression of candidate genes LIMK1 and ROCK1, which quere identified associated with copy number gain in 42.9% of vulvar carcinoma samples by CGH-array method, in order to determine better and more accurate prognostic values in vulvar carcinoma. Methods: 20 cases of vulvar cancer will be rescued from Hospital A. C. Camargo Biobank. LIMK1 and ROCK1 genes wil be validated by qRT-PCR and will be subjected to immunohistochemical tests in 150 samples of vulvar cancer. Functional analysis of these genes in cell culture SW954 (squamous cell carcinoma of the vulva) will be carried out by transfection with interference RNA (siRNA) method and viability of these cells will be determined by MTT assay. All data generated will be subjected to statistical tests correlating them with clinical data of patients intending to identify better prognostic factors. (AU)