Differential expression of CDC25 phosphatases splice variants in human breast cancer cells

Background: CDC25 phosphatases control cell cycle progression by activating cyclin dependent kinases. The three CDC25 isoforms encoding genes are submitted to alternative splicing events which generate at least two variants for CDC25A and five for both CDC25B and CDC25C. An over-expression of CDC25 was reported in several types of cancer, including breast cancer, and is often associated with a poor prognosis. Nevertheless, most of the previous studies did not address the expression of CDC25 splice variants. Here, we evaluated CDC25 spliced transcripts expression in anti-cancerous drug-sensitive and resistant breast cancer cell lines in order to identify potential breast cancer biomarkers. Methods: CDC25 splice variants mRNA levels were evaluated by semi-quantitative RT-PCR and by an original real-time RT-PCR assay. Results: CDC25 spliced transcripts are differentially expres-sed in the breast cancer cell lines studied. An up-regulation of CDC25A2 variant and an increase of the CDC25C5/C1 ratio are associated to the multidrug-resistance in VCREMS and DOXOR breast cancer cells, compared to their sensitive counterpart cell line MCF-7. Additionally, CDC25B2 tran-script is exclusively over-expressed in VCREMS resistant cells and could therefore be involved in the development of certain type of drug resistance. Conclusions: CDC25 splice variants could represent interesting potential breast cancer prognostic biomarkers.

eRF3a/GSPT1 12-GGC allele increases the susceptibility for breast cancer development

It is now widely recognized that translation factors are involved in cancer development and that components of the translation machinery that are deregulated in cancer cells may become targets for cancer therapy. The eukaryotic Release Factor 3 (eRF3) is a GTPase that associates with eRF1 in a complex that mediates translation termination. eRF3a/GSPT1 first exon contains a (GGC)n expansion coding for proteins with different N-terminal extremities. Herein we show that the longer allele (12-GGC) is present in 5.1% (7/137) of the breast cancer patients analysed and is absent in the control population (0/135), corresponding to an increased risk for cancer development, as revealed by Odds Ratio analysis. mRNA quantification suggests that patients with the 12-GGC allele overexpress eRF3a/GSPT1 in tumor tissues relative to the normal adjacent tissues. However, using an in vivo assay for translation termination in HEK293 cells, we do not detect any difference in the activity of the eRF3a proteins encoded by the various eRF3a/GSPT1 alleles. Although the connection between the presence of eRF3a/GSPT1 12-GGC allele and tumorigenesis is still unknown, our data suggest that the presence of the 12-GGC allele provides a potential novel risk marker for various types of cancer.

Modulation of translation factor's gene expression by histone deacetylase inhibitors in breast cancer cells

The histone deacetylase inhibitors sodium butyrate (NaBu) and trichostatin A (TSA) exhibit anti-proliferative activity by causing cell cycle arrest and apoptosis. The mechanisms by which NaBu and TSA cause apoptosis and cell cycle arrest are not yet completely clarified, although these agents are known to modulate the expression of several genes including cell-cycle- and apoptosis-related genes. The enzymes involved in the process of translation have important roles in controlling cell growth and apoptosis, and several of these translation factors have been described as having a causal role in the development of cancer. The expression patterns of the translation mechanism, namely of the elongation factors eEF1A1 and eEF1A2, and of the termination factors eRF1 and eRF3, were studied in the breast cancer cell line MCF-7 by real-time quantitative reverse transcription-polymerase chain reaction after a 24-h treatment with NaBu and TSA. NaBu induced inhibition of translation factors' transcription, whereas TSA caused an increase in mRNA levels. Thus, these two agents may modulate the expression of translation factors through different pathways. We propose that the inhibition caused by NaBu may, in part, be responsible for the cell cycle arrest and apoptosis induced by this agent in MCF-7 cells.

Avaliação da caracterização de lesões em mamografia com recurso a sistemas CAD (Computer-Aided Diagnosis)

Os sistemas Computer-Aided Diagnosis (CAD) auxiliam a deteção e diferenciação de lesões benignas e malignas, aumentando a performance no diagnóstico do cancro da mama. As lesões da mama estão fortemente correlacionadas com a forma do contorno: lesões benignas apresentam contornos regulares, enquanto as lesões malignas tendem a apresentar contornos irregulares. Desta forma, a utilização de medidas quantitativas, como a dimensão fractal (DF), pode ajudar na caracterização dos contornos regulares ou irregulares de uma lesão. O principal objetivo deste estudo é verificar se a utilização concomitante de 2 (ou mais) medidas de DF – uma tradicionalmente utilizada, a qual foi designada por “DF de contorno”; outra proposta por nós, designada por “DF de área” – e ainda 3 medidas obtidas a partir destas, por operações de dilatação/erosão e por normalização de uma das medidas anteriores, melhoram a capacidade de caracterização de acordo com a escala BIRADS (Breast Imaging Reporting and Data System) e o tipo de lesão. As medidas de DF (DF contorno e DF área) foram calculadas através da aplicação do método box-counting, diretamente em imagens de lesões segmentadas e após a aplicação de um algoritmo de dilatação/erosão. A última medida baseia-se na diferença normalizada entre as duas medidas DF de área antes e após a aplicação do algoritmo de dilatação/erosão. Os resultados demonstram que a medida DF de contorno é uma ferramenta útil na diferenciação de lesões, de acordo com a escala BIRADS e o tipo de lesão; no entanto, em algumas situações, ocorrem alguns erros. O uso combinado desta medida com as quatro medidas propostas pode melhorar a classificação das lesões.