Unmasking epithelial-mesenchymal transition in a breast cancer primary culture: a study report

Background: Immortalized cancer cell lines are now well-established procedures in biomedicine for a morecomplete understanding of cellular processes in cancer. However, they are more useful in preparation of freshtumour tissue, in order to obtain cancer cells with highly preserved individual tumour properties. In the presentstudy we report an analytical investigation on a breast cancer primary cell culture isolated from a surgical specimenobtained from a patient with an infiltrating ductal carcinoma. The objective of the research was to revealunrecognized aspects of neoplastic cells, typical of the tumour from where the cells were derived, but masked infixed tissue sections, in order to better predict the aggressive potentiality of the tumour.Findings: Using a combination of mechanical and enzymatic treatment, the tumour tissue was dissociatedimmediately after surgical removal. The primary cells were isolated by differential cell centrifugation and grown inselective media. Immunocytochemistry and quantitative RT-PCR analysis were performed to detect the presence ofspecific biomarkers at protein and transcript level.The isolated primary breast cancer cells displayed phenotypic behaviour, characteristic of malignant cells andexpression of several mesenchymal markers, revealing a strong signature for the epithelial-to-mesenchymaltransition associated to a stellate morphology with a number of cellular protrusions and the attitude to overgrowas multilayered overlapping cellular foci.Conclusions: Our data are a further meaningful indication that primary cell cultures represent a powerful systemthat could be applied to those cases deserving a deeper investigation at molecular level in order to designindividualized anticancer therapies in the future.