High-throughput time-resolved FRET reveals Akt/PKB activation as a poor prognostic marker in breast cancer
Autor: | Pierre Leboucher, Tamara Bunting, Gordon Stamp, Emma Nye, Nirmal Jethwa, Banafshé Larijani, Stefanie S. Jeffrey, Véronique Calleja, Richard Stone, Julien de Naurois, Selvaraju Veeriah, Peter J. Parker |
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Rok vydání: | 2014 |
Předmět: |
Cancer Research
Poor prognosis Tissue microarray Cancer Breast Neoplasms Biology medicine.disease Prognosis Molecular biology Disease-Free Survival High-Throughput Screening Assays Breast cancer Förster resonance energy transfer Oncology Cell Line Tumor medicine Cancer research Biomarkers Tumor Fluorescence Resonance Energy Transfer Biomarker (medicine) Humans Female DNA microarray Protein kinase B Proto-Oncogene Proteins c-akt |
Zdroj: | Cancer research. 74(18) |
ISSN: | 1538-7445 |
Popis: | Dysregulation of the Akt/PKB pathway has been associated with poor prognosis in several human carcinomas. Current approaches to assess Akt activation rely on intensity-based methods, which are limited by the subjectivity of manual scoring and poor specificity. Here, we report the development of a novel assay using amplified, time-resolved Förster resonance energy transfer (FRET), which is highly specific and sensitive and can be adapted to any protein. Using this approach to analyze primary breast tissue microarrays, we quantified levels of activated pAkt at a spatial resolution that revealed molecular heterogeneity within tumors. High pAkt status assessed by amplified FRET correlated with worse disease-free survival. Our findings support the use of amplified FRET to determine pAkt status in cancer tissues as candidate biomarker for the identification of high-risk patients. Cancer Res; 74(18); 4983–95. ©2014 AACR. |
Databáze: | OpenAIRE |
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