Identificação do gene EBER1 e EBNA1 do vírus de Epstein Barr (EBV) em tecidos de pacientes com doença de Hodgkin na região Norte do Brasil / Identification of gene EBER1 and EBNA 1 of Epstein Barr virus (EBV) in tissues of patients with Hodgkin’s disease in Northern Brazil

O vírus de Epstein Barr (EBV) é o agente causador da mononucle¬ose infecciosa e está associado a várias desordens proliferativas malignas tais como: linfoma de Burkitt, linfoma de Hodgkin e lin¬fomas não Hodgkin. Objetivo: detectar o genoma do EBV mediante a identificação dos genes EBER1 e EBNA1 em casos de doença de Hodgkin. Métodos: um total de 65 casos de linfomas diagnosti¬cados no Hospital Ophir Loyola no período de 1996 e 2005 foram analisados no Instituto Evandro Chagas, Ananindeua, Brasil. Todos os espécimes parafinizados foram analisados por hibridização in situ (gene EBER1) e PCR em tempo real (EBNA1). Resultados: do total, 64,6% (42/65) dos pacientes eram do sexo masculino e 35,4% (23/65) do sexo feminino. O EBV foi identificado por HIS nas células Reed Sternberg e variantes em 76,9% (50/65) dos casos com idade média de 28,3 anos (variação 2-84 anos). Os subtipos histológicos de casos EBV-positivos foram os seguintes: esclerose nodular em 50% (25/50), celularidade mista em 28% (14/50), depleção linfocitária em 14% (7/50) e predominância linfocitária em 8% (4/50). O DNA do EBV foi detectado em 53% (26/49) dos casos de doença de Hodgkin com um coeficiente de regressão para a curva padrão de 0,99. Conclusão: este estudo foi a primeira descrição do vírus de Epstein Barr em casos de linfoma de Hodgkin na Amazônia Brasileira, reforçando a hipótese de que o EBV seja um co-fator no processo de transformação neoplásica em conjunto com a predisposição genética e imunidade do paciente

Introduction: EBV is the causative agent of infectious mononucleosis and is associated with several malignant proliferative disorders such as Burkitt’s lymphoma, Hodgkin’s lymphoma, some B and T cell non-Hodgkin’s lymphomas. Objective: The main objective of the study was to determine the prevalence of EBER 1 gene and EBNA1 gene in cases of Hodgkin’s disease. Material and Methods: A total of 65 cases of lymphomas diagnosed between 1996 and 2005 were obtained from “Instituto Ofir Loyola” and analyzed at the “Instituto Evandro Chagas” Ananindeua, Brazil. The EBV antigens using EBER 1 probe in situ hybridization (HIS) and real time quantitative PCR. Results: From the total obtained, 64.6% (42/65) were male and 35.4% (23/65) female. EBV was identified in the Reed- Sternberg cells and variants in 76.9% (50/65) of Hodgkin’s disease cases, the median age were 28.3 years (range 2-84). The histologic subtypes of EBV-positive cases were as follows: nodular sclerosis in 50% (25/50), mixed cellularity in 28% (14/50), lymphocyte depletion in 14% (7/50) and lymphocyte predominance in 8% (4/50). We detected EBV DNA in 53% (26/49) with a coefficient of regression for the standard curve of a minimum of 0.99. Conclusion: These results were the first demonstration of the role of Epstein Barr virus in cases of Hodgkin diseases in northern Brazil and are consistent with the hypothesis that the presence of EBV during neoplasic transformation could be an additional cofactor acting together with both genetic predisposition and immunity of the patient

Adenovirus - mediated Herpes Simplex Virus Thymidine Kinase Gene Transference and Combined Drug Therapy Leads to Opoptosis of Human Epithelial Ovarian Cancer Cells / 中华妇产科杂志

Objective To evaluate the treatment and apoptosis effect of adenovirus-mediated herpes simplex virus thymidine kinase (HSV 1-tk) gene transference followed by administration of ganciclovir (GCV) and acyclovir (ACV) on ovarian epithelial cancer cells. Methods Recombinant adenovirus was amplificated and purified by routine method. The expression of HSV 1-tk gene was assayed by polymerase chain reaction (PCR). The efficiency of recombinant Advtk transference was evaluated. The cytotoxicity efficacy of TYK cells that carry HSV 1-tk gene was evaluated followed by GCV、ACV administration after the transference of Advtk. The changes and apoptosis of TYK cells that carry HSV 1-tk gene were observed by means of analysis of DNA fragmentation and electronic microscopy. Results Adenovirus was amplificated and purified in large amount. PCR assay showed 404 bp special band. When the multiplicities of infection (MOI) was 100, the transduction rate was 98 9%. The inhibition rates of TYK cells that carry HSV 1-tk gene increased with the increase of MOI when the same concentration of GCV、 ACV were given. When the MOI was same, the inhibition rates were also increased with the increase concentration of GCV、ACV. Apoptosis of TYK cells that carry HSV 1-tk gene after administration of GCV was observed by means of analysis of DNA fragmentation and electronic microscopy. Conclusions The HSV 1-tk gene can effectively transfered into TYK cells by recombinant replicated-deficient adenovirus vector, GCV、ACV can effectively kill TYK cells that carry HSV 1-tk gene in vitro. Apoptosis may be the mechanism of the killing effect.