Investigação da integração genômica (método IntG) para determinação direta dos padrões dos status físico do genoma do papilomavírus humano em relação ao genoma das células infectadas

Abstract

The immortalization and transformation power of neoplastic cells are due to the viral oncogenes E6 and E7, which generate oncoproteins capable of mediating mitogenic and antiapoptotic stimuli through their interaction with cell cycle regulatory proteins. The HPV genome is maintained in episomal form in the early stage of low-grade dysplastic lesions, however, in advanced precancerous cells and most carcinomas, it is integrated into the DNA of the infected cell. This integration causes increased expression of the E6 and E7 oncogenes, promoting selective advantages for tumor growth. This dissertation thesis refers to the areas of molecular biology and in vitro molecular diagnostics. It consists of directly determining the physical status of the HPV genome in relation to the genome of infected cells, which can develop neoplastic processes, such as Cervical Cancer – CC. The methodology used in the work, called IntG-HPV, was validated on DNA samples from the CCU cell lines SiHa, Ca Ski and uterine cervix desquamation cells with low-grade lesions positive for HPV-16, by PCR reaction. Multiplex. Comparative analysis using Fisher's exact test showed a statistically significant difference p-value > 0.05. Thus, the results suggest the presence of an episomal pattern in the aforementioned desquamation cells, a mixed pattern in the Ca Ski cell line and an integrated pattern in the SiHa cell line. Therefore, the IntG-HPV method innovates in the direct analysis of the integration of the HPV genome into the genome of the infected cell, being a molecular diagnostic and prognostic test for screening and prevention of CC.