Potencial de transformação maligna de células B-1 resistentes à radiação ionizante
Data
2015-09-30
Autores
Guimaraes, Caroline Ferreira 
Orientadores
Popi, Ana Flavia
Tipo
Dissertação de mestrado
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Resumo
Analisar o potencial de malignidade de células B-1 resistentes à radiação ionizante in vitro e verificar a capacidade destas células sobreviverem e proliferarem após múltiplas transferências em camundongos imunocomprometidos NOD/SCID gamma null. Métodos: Células provenientes da cavidade peritoneal foram purificadas para obtenção de células B-1. Estas células foram submetidas à radiação ionizante e mantidas em co-cultivo com células OP9 por 3, 7 e 15 dias. Após esses períodos foram analisados os seguintes parâmetros: viabilidade, proliferação, morfologia, expressão de genes da família do Bcl, hiperploidia e indução a apoptose com estímulo de anti-IgM. Células B-1 obtidas de camundongos transgênicos para GFP foram purificadas, irradiadas e injetadas intraperitonealmente em camundongos imunocomprometidos. A presença de células B-1 GFP+ em diversos órgãos destes animais foi analisada após sucessivas passagens. Resultados: Células B-1 radiorresistentes demonstraram sobrevida após longos períodos de cultura, com aumento na proliferação em relação a células B-1 não irradiadas (controle). Além disso, células B-1 radiorresistentes apresentaram hiperplodia, alterações orfológicas, aumento da indução da apoptose com estímulo de anti-IgM. Também foi observado que estas células foram capazes de sobreviver e expandir in vivo, sendo detectadas em fígado, baço, medula óssea e cavidade peritoneal de camundongos imunocomprometidos. Conclusão: Células B-1 radiorresistentes apresentam modificações que sugerem aquisição de potencial maligno
This study aims to investigate the malignant potential of ionizing-radiation resistant B-1 cells in vitro and evaluate the ability of these cells survive and proliferate in vivo. Methods: Purified peritoneal B-1 cells were submitted to ionizing radiation and maintained in co-culture with OP9 cells for 3, 7 and 15 days. Viability, proliferation, morphology, Bcl family gene expression, DNA quantity and induction of apoptosis with anti-IgM stimulation were analysed in these cells. To analyse the capacity of radioresistant B-1 cells to survive and expand in vivo, B-1 cells were obtained from transgenic mice to GFP. B-1 GFP+ cells were irradiated and injected intraperitoneally in immunodeficient mice NOD/SCID gamma null. The presence of B-1 GFP+ cells in several organs of animals was analyzed after successive passages. Results: Radioresistant B-1 cells were able to survive long periods in culture. Further, these cells show proliferation index increase in relation to non irradiated B-1 cells (control). In addition, radioresistant B-1 cells showed hyperploid, morphologic alterations, increased induction of apoptosis after anti-IgM stimulation. It was also observed that these cells were able to survive and expand in vivo. These cells were detected in liver, spleen, bone marrow and peritoneal cavity of immunodeficient mice. Conclusion: Radioresistant B-1 cells showed modifications in some characteristics that could be related to induction of malignant potential.
This study aims to investigate the malignant potential of ionizing-radiation resistant B-1 cells in vitro and evaluate the ability of these cells survive and proliferate in vivo. Methods: Purified peritoneal B-1 cells were submitted to ionizing radiation and maintained in co-culture with OP9 cells for 3, 7 and 15 days. Viability, proliferation, morphology, Bcl family gene expression, DNA quantity and induction of apoptosis with anti-IgM stimulation were analysed in these cells. To analyse the capacity of radioresistant B-1 cells to survive and expand in vivo, B-1 cells were obtained from transgenic mice to GFP. B-1 GFP+ cells were irradiated and injected intraperitoneally in immunodeficient mice NOD/SCID gamma null. The presence of B-1 GFP+ cells in several organs of animals was analyzed after successive passages. Results: Radioresistant B-1 cells were able to survive long periods in culture. Further, these cells show proliferation index increase in relation to non irradiated B-1 cells (control). In addition, radioresistant B-1 cells showed hyperploid, morphologic alterations, increased induction of apoptosis after anti-IgM stimulation. It was also observed that these cells were able to survive and expand in vivo. These cells were detected in liver, spleen, bone marrow and peritoneal cavity of immunodeficient mice. Conclusion: Radioresistant B-1 cells showed modifications in some characteristics that could be related to induction of malignant potential.
Descrição
Citação
GUIMARAES, Caroline Ferreira. Potencial de transformação maligna de células B-1 resistentes à radiação ionizante. 2015. 55 f. Dissertação (Mestrado em Microbiologia e Imunologia) - Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, 2015.