title: Unmasking epithelial-mesenchymal transition in a breast cancer primary culture: a study report.
abstract: BACKGROUND
  Immortalized cancer cell lines are now well-established procedures in biomedicine for a more complete understanding of cellular processes in cancer. However, they are more useful in preparation of fresh tumour tissue, in order to obtain cancer cells with highly preserved individual tumour properties. In the present study we report an analytical investigation on a breast cancer primary cell culture isolated from a surgical specimen obtained from a patient with an infiltrating ductal carcinoma. The objective of the research was to reveal unrecognized aspects of neoplastic cells, typical of the tumour from where the cells were derived, but masked in fixed tissue sections, in order to better predict the aggressive potentiality of the tumour.
  
  
  RESULTS
  Using a combination of mechanical and enzymatic treatment, the tumour tissue was dissociated immediately after surgical removal. The primary cells were isolated by differential cell centrifugation and grown in selective media. Immunocytochemistry and quantitative RT-PCR analysis were performed to detect the presence of specific biomarkers at protein and transcript level.The isolated primary breast cancer cells displayed phenotypic behaviour, characteristic of malignant cells and expression of several mesenchymal markers, revealing a strong signature for the epithelial-to-mesenchymal transition associated to a stellate morphology with a number of cellular protrusions and the attitude to overgrow as multilayered overlapping cellular foci.
  
  
  CONCLUSIONS
  Our data are a further meaningful indication that primary cell cultures represent a powerful system that could be applied to those cases deserving a deeper investigation at molecular level in order to design individualized anticancer therapies in the future.
authors: Institute of Molecular Bioimaging and Physiology (IBFM), National Council of Researches (CNR), Cefalù-Segrate, Italy. luigi.minafra@polooncologicocefalu.it
authors: Luigi
authors: L
authors: Minafra
authors: Rossana
authors: R
authors: Norata
authors: Valentina
authors: V
authors: Bravatà
authors: Massimo
authors: M
authors: Viola
authors: Carmelo
authors: C
authors: Lupo
authors: Cecilia
authors: C
authors: Gelfi
authors: Cristina
authors: C
authors: Messa
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D018270:Carcinoma, Ductal, Breast / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000235:genetics / Q000378:metabolism; D008858:Microscopy, Phase-Contrast; D009130:Muscle, Smooth / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D014408:Biomarkers, Tumor; D015820:Cadherins; D014746:Vimentin; D007633:Keratins