Invasiveness of breast carcinoma cells and transcript profile: Eph receptors and ephrin ligands as molecular markers of potential diagnostic and prognostic application
Section snippets
Materials and methods
Cell cultures. All cells were cultured at 37 °C/7% CO2. MCF-10A cells were grown in 1:1 DMEM:F12 media (Gibco) with 5% Horse Serum (Gibco), 20 mM Hepes, 10 ng/ml EGF (Invitrogen), 10 ml/L PenStrep-Glutamine 10,000 U/ml penicillin, 10,000 μg/ml streptomycin, and 29.2 mg/ml l-glutamine (Gibco), 10 g/ml insulin (Invitrogen), 0.1 μg/ml Cholera Toxin (Sigma), and 500 ng/ml Hydrocortisone (Sigma). MCF-7 and MDA-MB-231 cells were grown in DMEM (Gibco) supplemented with 2 mM l-glutamine (Gibco), 1 mM sodium
Results
We have first confirmed the invasive phenotype of the breast carcinoma cell lines used in this study. We have then analyzed the abundance of 13 members of the Eph receptor family and 8 members of the ephrin ligand family in three cell lines, namely, MCF-10A, MCF-7, and MDA-MB-231. The results are presented in the following manner: (1) The relative abundance of each Eph receptor and ephrin ligand in individual cell lines to obtain a cell-type profile of these transcripts. (2) A comparative
Discussion
The evaluation of cell lines established from normal and cancer tissue has often served as a primer for biological characterization of a number of genes involved in tumorigenesis. While global gene profiling via gene microarrays is currently being used to arrive at a molecular signature of tumor tissues [42], [43], the analysis of such large-scale expression profiles is a major challenge. We reasoned that a systematic analysis of gene families involved in specific signaling pathways may allow
Acknowledgements
We wish to acknowledge the Department of Defense for supporting the P.I’s (R.P.K.) research. Our thanks to Dr. Berish Rubin and Dr. Sylvia Anderson for use of real time PCR machine, Jinsong Qiu for his help with designing and running real time experiments, and Nagaraja Ganachari for technical assistance.
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2020, ToxiconCitation Excerpt :Finally, the expression levels of EphB6 in cancer cells has been linked to their progression and development, with losses in EphB6 expression often indicative of a more aggressive cancer type. Downregulations in EphB6 receptor have been seen in breast cancer cells, where it has been found to interact with c-Cbl to function as a tumour suppressor and to prevent tumour cell invasiveness (Fox & Kandpal, 2004, 2006, 2009; Truitt et al., 2010). EphB6 expression has been shown to partially supress epithelial-to-mesenchymal transition in triple negative breast cancer cells and to reduce tumour drug-resistance to DNA-damaging drugs, resulting in better chances for recurrence-free survival in patients with higher EphB6 expressing tumours (Toosi et al., 2018).
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2018, Cancer LettersCitation Excerpt :It subsequently investigates the system's role in tumor microenvironment, then its clinical significance with a focus on the IHC-based groups, grading, staging and survival, and finishes with a discussion of the findings, as well as some future perspectives. EphA2 is upregulated in the MDA-MB-231 cell line and activated primarily through ligand-independent signaling [20,91,92]. However, ephrin-A1 stimulated EphA2 forward signaling suppresses EphA2 when tested in the same TNBC cell line, it also induces apoptosis [48,93].