Padronização dos ensaios para caracterização da população celular presente no líquido ascítico de pacientes com câncer epitelial de ovário atendidas no serviço de ginecologia oncológica do Hospital das Clínicas/UFMG

Abstract

Ovarian cancer (OC) is the fifth cause of cancer deaths in North America. In Brazil, 6,150 new cases are estimated for this year according to INCA. One of the difficulties in OC treatment is the refractory disease, wherein the patients present disease progression and ascites. Ascites is associate to intraperitoneal metastasis and/or distant metastasis as results conditions patients normally have bad clinical outcome and poor prognosis. This study aimed standardize methods for isolate, culture and characterize subpopulations present in ascites from Epithelial Ovarian Cancer patients, as study model of tumor proliferation and resistance mechanisms. The samples of ascitic fluid were collected from three patients attended at the Gynecology Oncology Service of Hospital das Clínicas/UFMG (HC-UFMG). The primary cell culture was established, with confluent grow in fifth day. Many assays were done to attempt cell characterization. The immunofluorescence assay evaluated the E/P cadherin presence. The results showed negative expression of these molecules, which could mean passage of the cells through the epithelial-mesenchymal (EMT) transition process, phenomenon that contributes to the metastatic phenotype, or indicate fibroblast presence. In addition, the cytoskeleton and cell nucleus were marked to verify morphology preservation of these structures, which revealed an irregular cell nucleus and cytoskeleton and binuclear cells. The expression of imunomarkers related to the mesenchymal epithelial transition pathways and pluripotency in ascites samples of CEO patients by flow cytometry could not be done, because procedures and counting necessaries to this assay were prevented by formation of cell pellets. Studies are in progress to optimize assays for phenotype and imunophenotype characterization of this type of sample.