Patrícia Dias Fernandes1, Fabiana S Guerra2, Natália M Sales2, Thais B Sardella2, Sonia Jancar3, Josiane S Neves4. 1. Universidade Federal do Rio de Janeiro, Instituto de Ciências Biomédicas, Laboratório de Farmacologia da Dor e da Inflamação, Brazil. Electronic address: patricia.dias@icb.ufrj.br. 2. Universidade Federal do Rio de Janeiro, Instituto de Ciências Biomédicas, Laboratório de Farmacologia da Dor e da Inflamação, Brazil. 3. Universidade de São Paulo, Instituto de Ciências Biomédicas, Departamento de Imunologia, Universidade de São Paulo, Brazil. 4. Universidade Federal do Rio de Janeiro, Instituto de Ciências Biomédicas, Laboratório Compartilhado, Brazil.
Abstract
INTRODUCTION: Ehrlich tumor is a mammary adenocarcinoma with aggressive behavior. Inoculated in mice peritoneal cavity, the Ehrlich tumor grows in ascitic form (EAT). Since inflammation modulates tumor progression we further investigated the inflammatory response during EAT growth. METHODS: Balb/C mice were intraperitoneal inoculated with 5×10(5) Ehrlich cells and after every 2days, blood samples were collected for hemoglobin, hematocrit, platelets and leukocytes counts. The ascitic fluid was collected for protein concentration and cell count. Phenotype analysis of the peritoneal cells was made by FACS, prostaglandin E2 (PGE2) and cytokines by ELISA, nitric oxide (NO) by nitrate conversion protocol, and cyclooxygenase-1 (COX1), COX2 and inducible nitric oxide synthase (iNOS) by immunoblotting. RESULTS: Following EAT inoculation into the peritoneal cavity there was a rapid increase in ascitis volume and protein concentration. The cell number in ascitis remained stable until day 8 (lag phase) followed by a sharp increase. As tumor progressed, blood leukocytes increased and erythrocyte decreased. Phenotypic analysis showed that during the lag phase the percentage of F4/80(+) cells remained similar to control levels and around 7% of this population was also positive for the GR1 marker. These double-positive cells (probably inflammatory monocytes) markedly increased at day 6. The percentage of F4/80-GR1(+)cells (probably neutrophils) was low and did not significantly vary during tumor progression. CD4(+) and CD8(+) cells were not detected in the time points analyzed. iNOS and COX1 expression increased after day 2 reaching peak levels on day 10. COX2 enzyme expression did not change significantly over time. Sustained increase in PGE2 and NO levels was observed. IL-10 and MCP-1 peaked at day 14 and IL-1β increased progressively till day 10. IFN-γ levels were low till day 10, increasing progressively after that. DISCUSSION: These data extended the characterization of the inflammatory response during Ehrlich ascitis tumor growth, further validating it as a useful model for antitumor drugs screening.
INTRODUCTION:Ehrlich tumor is a mammary adenocarcinoma with aggressive behavior. Inoculated in mice peritoneal cavity, the Ehrlich tumor grows in ascitic form (EAT). Since inflammation modulates tumor progression we further investigated the inflammatory response during EAT growth. METHODS: Balb/C mice were intraperitoneal inoculated with 5×10(5) Ehrlich cells and after every 2days, blood samples were collected for hemoglobin, hematocrit, platelets and leukocytes counts. The ascitic fluid was collected for protein concentration and cell count. Phenotype analysis of the peritoneal cells was made by FACS, prostaglandin E2 (PGE2) and cytokines by ELISA, nitric oxide (NO) by nitrate conversion protocol, and cyclooxygenase-1 (COX1), COX2 and inducible nitric oxide synthase (iNOS) by immunoblotting. RESULTS: Following EAT inoculation into the peritoneal cavity there was a rapid increase in ascitis volume and protein concentration. The cell number in ascitis remained stable until day 8 (lag phase) followed by a sharp increase. As tumor progressed, blood leukocytes increased and erythrocyte decreased. Phenotypic analysis showed that during the lag phase the percentage of F4/80(+) cells remained similar to control levels and around 7% of this population was also positive for the GR1 marker. These double-positive cells (probably inflammatory monocytes) markedly increased at day 6. The percentage of F4/80-GR1(+)cells (probably neutrophils) was low and did not significantly vary during tumor progression. CD4(+) and CD8(+) cells were not detected in the time points analyzed. iNOS and COX1 expression increased after day 2 reaching peak levels on day 10. COX2 enzyme expression did not change significantly over time. Sustained increase in PGE2 and NO levels was observed. IL-10 and MCP-1 peaked at day 14 and IL-1β increased progressively till day 10. IFN-γ levels were low till day 10, increasing progressively after that. DISCUSSION: These data extended the characterization of the inflammatory response during Ehrlich ascitis tumor growth, further validating it as a useful model for antitumor drugs screening.
Authors: Cassia Calixto-Campos; Mab P Corrêa; Thacyana T Carvalho; Ana C Zarpelon; Miriam S N Hohmann; Ana C Rossaneis; Leticia Coelho-Silva; Wander R Pavanelli; Phileno Pinge-Filho; Jefferson Crespigio; Catia C F Bernardy; Rubia Casagrande; Waldiceu A Verri Journal: Anal Cell Pathol (Amst) Date: 2015-08-13 Impact factor: 2.916
Authors: Ed Carlos Rey Moura; Plinio da Cunha Leal; Izabel Cristina Portela Bogéa Serra; Bruno de Paulo Ribeiro; Johnny Ramos do Nascimento; Flavia Raquel Fernandes do Nascimento; Rioko Kimiko Sakata Journal: BMC Res Notes Date: 2018-07-31
Authors: Plinio da Cunha Leal; Ed Carlos Rey Moura; Rachel Jorge Dino Cossetti; Johnny Ramos do Nascimento; Izabel Cristina Portela Bogéa Serra; Bruno de Paulo Ribeiro; Andre Álvares Marques Vale; Ana Paula Silva de Azevedo Dos Santos; Flavia Raquel Fernandes do Nascimento; Rioko Kimiko Sakata Journal: BMC Res Notes Date: 2019-01-25