Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom. Centre Emotion, Hôpital de la Pitié Salpetriere, Paris, France.
Cancer Res. 2014 Sep 15;74(18):4983-95. doi: 10.1158/0008-5472.CAN-13-3382. Epub 2014 Jun 26.
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.
Akt/PKB 通路的失调与几种人类癌的不良预后相关。目前评估 Akt 激活的方法依赖于基于强度的方法，该方法受到手动评分的主观性和特异性差的限制。在此，我们报告了一种使用扩增的、时间分辨的Förster 共振能量转移（FRET）的新型测定方法的开发，该方法高度特异和敏感，并且可以适用于任何蛋白质。使用这种方法分析原发性乳腺组织微阵列，我们以揭示肿瘤内分子异质性的空间分辨率定量分析了激活的 pAkt 的水平。通过扩增的 FRET 评估的高 pAkt 状态与无病生存率差相关。我们的发现支持使用扩增的 FRET 来确定癌症组织中的 pAkt 状态，作为鉴定高危患者的候选生物标志物。
