Browsing by Author "Costa, Vera L."
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- Evaluation of immunoexpression and MDR1 promoter methylation levels in prostatic tissue samplesPublication . Moura, Inês; Costa, Vera L.; Pais, Irene; Ribeiro, Franclim R.; Henrique, Rui; Jerónimo, CarmenOs objectivos deste estudo foram: (1) Determinar os níveis de metilação do promotor do MDR1 em tecido prostático com adenocarcinoma (CaP), neoplasia intraepitelial prostática de alto grau (HGPIN), hiperplasia benigna (BPH) e tecido morfologicamente normal (MNP). (2) Correlacionar os níveis de metilação com a imunoexpressão da gp-P. Os nossos resultados demonstram que a hipermetilação do MDR1 constitui um mecanismo eficaz de regulação da sua expressão. Estudos futuros permitirão avaliar o impacto destes resultados na terapêutica do cancro da próstata. Our aims were: (1) To determine the MDR1 methylation levels in tissue of Prostate cancer (PCa), high grade prostatic intraepithelial neoplasia (HGPIN), benign prostatic hyperplasia (BPH) and morphologically normal prostate tissue (MNP); (2) to correlate the methylation levels of the MDR1 promoter with the immunoexpression of P-gp. Our results demonstrate that MDR1 hypermethylation constitutes an effective mechanism of P-gp expression regulation. Future studies will be able to evaluate the impact of these results in the treatment of PCa patients.
- A rapid and simple procedure for the establishment of human normal and cancer renal primary cell cultures from surgical specimensPublication . Valente, Maria João; Henrique, Rui; Costa, Vera L.; Jerónimo, Carmen; Carvalho, Félix; Bastos, Maria L.; Guedes de Pinho, Paula; Carvalho, MárciaThe kidney is a target organ for the toxicity of several xenobiotics and is also highly susceptible to the development of malignant tumors. In both cases, in vitro studies provide insight to cellular damage, and represent adequate models to study either the mechanisms underlying the toxic effects of several nephrotoxicants or therapeutic approaches in renal cancer. The development of efficient methods for the establishment of human normal and tumor renal cell models is hence crucial. In this study, a technically simple and rapid protocol for the isolation and culture of human proximal tubular epithelial cells and human renal tumor cells from surgical specimens is presented. Tumor and normal tissues were processed by using the same methodology, based on mechanical disaggregation of tissue followed by enzymatic digestion and cell purification by sequential sieving. The overall procedure takes roughly one hour. The resulting cell preparations have excellent viabilities and yield. Establishment of primary cultures from all specimens was achieved successfully. The origin of primary cultured cells was established through morphological evaluation. Normal cells purity was confirmed by immunofluorescent staining and reverse transcription-polymerase chain reaction analysis for expression of specific markers.