IBMC Institute of Molecular and Cell Biology

Porto, Portugal

IBMC Institute of Molecular and Cell Biology

Porto, Portugal
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da Cruz Paula A.,University of Porto | da Cruz Paula A.,Portuguese Oncology Institute IPO | Leitao C.,University of Porto | Leitao C.,IBMC Institute of Molecular and Cell Biology | And 14 more authors.
Virchows Archiv | Year: 2017

Breast cancer epithelial cells with the CD44+/CD24−/low phenotype possess tumor-initiating cells and epithelial-mesenchymal transition (EMT) capacity. Massive parallel sequencing can be an interesting approach to deepen the molecular characterization of these cells. We characterized CD44+/CD24−/cytokeratin(Ck)+/CD45− cells isolated through flow cytometry from 43 biopsy and 6 mastectomy samples harboring different benign and malignant breast lesions. The Ion Torrent Ampliseq Cancer Hotspot panel v2 (CHPv2) was used for the identification of somatic mutations in the DNA extracted from isolated CD44+/CD24−/Ck+/CD45− cells. E-Cadherin and vimentin immunohistochemistry was performed on sections from the corresponding formalin-fixed, paraffin-embedded (FFPE) blocks. The percentage of CD44+/CD24−/Ck+/CD45− cells increased significantly from non-malignant to malignant lesions and in association with a significant increase in the expression of vimentin. Non-malignant lesions harbored only a single-nucleotide polymorphism (SNP). Mutations in the tumor suppressor p53 (TP53), NOTCH homolog 1 (NOTCH1), phosphatase and tensin homolog (PTEN), and v-akt murine thymoma viral oncogene homolog 1 (AKT1) genes were found in isolated CD44+/CD24−/Ck+/CD45− cells from ductal carcinomas in situ (DCIS). Additional mutations in the colony-stimulating factor 1 receptor (CSF1R), ret proto-oncogene (RET), and TP53 genes were also identified in invasive ductal carcinomas (IDCs). The use of massive parallel sequencing technology for this type of application revealed to be extremely effective even when using small amounts of DNA extracted from a low number of cells. Additional studies are now required using larger cohorts to design an appropriate mutational profile for this phenotype. © 2017 Springer-Verlag Berlin Heidelberg


Pereira P.,University of Minho | Pedrosa S.S.,University of Minho | Correia A.,IBMC Institute of Molecular and Cell Biology | Lima C.F.,University of Minho | And 5 more authors.
Toxicology in Vitro | Year: 2015

The research of chitosan-based nanogel for biomedical applications has grown exponentially in the last years; however, its biocompatibility is still insufficiently reported. Hence, the present work provides a thorough study of the biocompatibility of a glycol chitosan (GC) nanogel. The obtained results showed that GC nanogel induced slight decrease on metabolic activity of RAW, 3T3 and HMEC cell cultures, although no effect on cell membrane integrity was verified. The nanogel does not promote cell death by apoptosis and/or necrosis, exception made for the HMEC cell line challenged with the higher GC nanogel concentration. Cell cycle arrest on G1 phase was observed only in the case of RAW cells. Remarkably, the nanogel is poorly internalized by bone marrow derived macrophages and does not trigger the activation of the complement system. GC nanogel blood compatibility was confirmed through haemolysis and whole blood clotting time assays. Overall, the results demonstrated the safety of the use of the GC nanogel as drug delivery system. © 2014 Elsevier Ltd.


PubMed | University of Porto and IBMC Institute of Molecular and Cell Biology
Type: | Journal: Virchows Archiv : an international journal of pathology | Year: 2017

Breast cancer epithelial cells with the CD44

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