RESUMEN
BACKGROUND: The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression. METHODS: The study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells. RESULTS: We observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (ARID4A, CALR, GNB2L1, RNF10, SQSTM1, USP9X) were validated by real time PCR. CONCLUSIONS: A significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.
Asunto(s)
Regulación Neoplásica de la Expresión Génica , Neoplasias de la Boca/metabolismo , Proteoma/metabolismo , Anexina A5/metabolismo , Apoptosis , Proliferación Celular , Regulación hacia Abajo , Electroforesis en Gel Bidimensional , Fibroblastos/metabolismo , Genómica , Células Hep G2 , Humanos , Queratinas/metabolismo , Neoplasias de la Boca/genética , Hibridación de Ácido Nucleico , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Células del Estroma/metabolismo , Vimentina/metabolismoRESUMEN
Parâmetros baseados na probabilidade de origem do gene foram usados para descrever a variabilidade em uma população de búfalos da Embrapa Amazônia Oriental. A magnitude dos resultados foi de média a baixa (por volta de 20 animais), sugerindo que poucos fundadores contribuiriam para a formação da população. Dentre os 20 ancestrais que mais aportam genes aos machos - representando ao todo 71 por cento dos alelos -, 39 por cento, 26 por cento e 5 por cento, respectivamente, são as contribuições marginais das raças Murrah e Mediterrâneo e seus mestiços. Para as fêmeas, em que os 20 ancestrais aportam 67,5 por cento dos genes, 42 por cento e 26 por cento, respectivamente, são as contribuições marginais das raças Murrah e Mediterrâneo.
Parameters based on the probability of gene origin were used to describe genetic variability in a buffalo population from the Embrapa Amazônia Oriental, Belém, Pará, Brazil. The parameters generated medium to low values (around 20 animals) and suggested low founder representativeness. From the 20 ancestors that gave more genes to males (with 71 percent of alleles), genetic contributions were 39 percent, 26 percent, and 5 percent, respectively, for Murrah, Mediterraneo, and crossbreds. For females, these values were 42 percent and 26 percent for Murrah and Mediterraneo breeds.