We are analyzing https://link.springer.com/article/10.1186/bcr2896.

Title:
Epithelial to mesenchymal transition markers expressed in circulating tumour cells of early and metastatic breast cancer patients | Breast Cancer Research
Description:
Introduction Epithelial to mesenchymal transition (EMT) is considered an essential process in the metastatic cascade. EMT is characterised by upregulation of vimentin, Twist, Snail, Slug and Sip1 among others. Metastasis is also associated with the presence of circulating tumour cells (CTCs) and disseminated tumour cells in the blood and bone marrow, respectively, of breast cancer patients, but the expression of EMT markers in these cells has not been reported so far. Methods The expression of Twist and vimentin in CTCs of 25 metastatic and 25 early breast cancer patients was investigated by using double-immunofluorescence experiments in isolated peripheral blood mononuclear cell cytospins using anti-cytokeratin (anti-CK) anti-mouse (A45-B/B3) and anti-Twist or anti-vimentin anti-rabbit antibodies. Results Among early breast cancer patients, vimentin-and Twist-expressing CK+ CTCs were identified in 77% and 73% of the patients, respectively, and in 100% of the patients with metastatic breast cancer for both markers (P = 0.004 and P = 0.037, respectively). Among patients with early disease, 56% and 53% of the CK+ CTCs were double-stained with vimentin and Twist, and the corresponding values for metastatic patients were 74% and 97%, respectively (P = 0.005 and P = 0.0001, respectively). The median expression of CK+vimentin+ and CK+Twist+ cells per patient in metastatic patients was 98% and 100%, and in an adjuvant chemotherapy setting the corresponding numbers were 56% and 40.6%, respectively. Triple-staining experiments revealed that all CK+Twist+ or CK+vimentin+ cells were also CD45-, confirming their epithelial origin. Immunomagnetic separation of CTCs and triple-immunofluorescence with anti-CK/anti-Twist/anti-vimentin antibodies demonstrated that both mesenchymal markers could be coexpressed in the same CK+ cell, since 64% of the total identified CTCs were triple-stained. There was a significant correlation (P = 0.005) between the number of CTCs expressing Twist and vimentin within the same setting. Conclusions CTCs expressing Twist and vimentin, suggestive of EMT, are identified in patients with breast cancer. The high incidence of these cells in patients with metastatic disease compared to early stage breast cancer strongly supports the notion that EMT is involved in the metastatic potential of CTCs.
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Keywords {🔍}

cancer, cells, patients, breast, ctcs, twist, vimentin, expression, pubmed, article, metastatic, google, scholar, cas, early, cell, antibody, fluor, tumour, emt, circulating, antirabbit, blood, disease, experiments, stage, res, patient, figure, antimouse, original, tumor, epithelial, markers, antibodies, hela, mesenchymal, expressed, abb, transition, cytokeratin, system, panck, negative, samples, control, minutes, stained, alexa, clin,

Topics {✒️}

bone-marrow/peripheral-blood stem-cell products anti-ck/anti-twist/anti-vimentin antibodies demonstrated /b3 antibody/fitc anti-mouse anti-vimentin anti-rabbit antibodies /b3 anti-mouse antibody twist anti-mouse antibody /polyclonal vimentin anti-rabbit /polyclonal twist anti-rabbit anti-rabbit secondary antibodies double-immunofluorescence confocal laser-scanning anti-rabbit antibody anti-cd45 monoclonal antibody vimentin anti-rabbit antibodies anti-mouse antibody fitc-conjugated igg1 antibody vassilis georgoulias & sophia agelaki confocal laser-scanning microscope confocal laser-scanning microscopy twist anti-rabbit full size image anti-vimentin antibodies revealed epithelial-mesenchymal transition markers pan-ck/vimentin/cd45 antibodies article download pdf double-positive ctcs/total ctcs pi3k/akt signaling kinases hypoxia-inducible factor 1α hypoxia-inducible factor-1α ck-19 mrna-positive cells exclusively double-positive cells alexa fluor 633 alexa fluor 555 50 mg/ml penicillin/streptomycin hif-1α promotes metastasis real-time rt-pcr triple-staining experiments revealed anti-mouse stem cell ariol scanning microscopy detectable double-stained cells double-staining experiments revealed anti-pan-ck full access isotype antibody exclusively twist+ck+ cells performed triple-immunofluorescence experiments ariol system images abbreviations ariol system e-cadherin expression leads repressing e-cadherin expression

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         headline:Epithelial to mesenchymal transition markers expressed in circulating tumour cells of early and metastatic breast cancer patients
         description:Epithelial to mesenchymal transition (EMT) is considered an essential process in the metastatic cascade. EMT is characterised by upregulation of vimentin, Twist, Snail, Slug and Sip1 among others. Metastasis is also associated with the presence of circulating tumour cells (CTCs) and disseminated tumour cells in the blood and bone marrow, respectively, of breast cancer patients, but the expression of EMT markers in these cells has not been reported so far. The expression of Twist and vimentin in CTCs of 25 metastatic and 25 early breast cancer patients was investigated by using double-immunofluorescence experiments in isolated peripheral blood mononuclear cell cytospins using anti-cytokeratin (anti-CK) anti-mouse (A45-B/B3) and anti-Twist or anti-vimentin anti-rabbit antibodies. Among early breast cancer patients, vimentin-and Twist-expressing CK+ CTCs were identified in 77% and 73% of the patients, respectively, and in 100% of the patients with metastatic breast cancer for both markers (P = 0.004 and P = 0.037, respectively). Among patients with early disease, 56% and 53% of the CK+ CTCs were double-stained with vimentin and Twist, and the corresponding values for metastatic patients were 74% and 97%, respectively (P = 0.005 and P = 0.0001, respectively). The median expression of CK+vimentin+ and CK+Twist+ cells per patient in metastatic patients was 98% and 100%, and in an adjuvant chemotherapy setting the corresponding numbers were 56% and 40.6%, respectively. Triple-staining experiments revealed that all CK+Twist+ or CK+vimentin+ cells were also CD45-, confirming their epithelial origin. Immunomagnetic separation of CTCs and triple-immunofluorescence with anti-CK/anti-Twist/anti-vimentin antibodies demonstrated that both mesenchymal markers could be coexpressed in the same CK+ cell, since 64% of the total identified CTCs were triple-stained. There was a significant correlation (P = 0.005) between the number of CTCs expressing Twist and vimentin within the same setting. CTCs expressing Twist and vimentin, suggestive of EMT, are identified in patients with breast cancer. The high incidence of these cells in patients with metastatic disease compared to early stage breast cancer strongly supports the notion that EMT is involved in the metastatic potential of CTCs.
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      headline:Epithelial to mesenchymal transition markers expressed in circulating tumour cells of early and metastatic breast cancer patients
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