High-throughput time-resolved FRET reveals Akt/PKB activation as a poor prognostic marker in breast cancer.
High-throughput time-resolved FRET reveals Akt/PKB activation as a poor prognostic marker in breast cancer.
复制标题
高通量时间分辨 FRET 揭示 Akt/PKB 激活是乳腺癌的不良预后标志物。
DOI：
10.1158/0008-5472.can-13-3382
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发表时间：
2014
期刊：
影响因子：
11.2
通讯作者：
Larijani,Banafshé
中科院分区：
文献类型：
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作者：
Veeriah,Selvaraju;Leboucher,Pierre;deNaurois,Julien;Jethwa,Nirmal;Nye,Emma;Bunting,Tamara;Stone,Richard;Stamp,Gordon;Calleja,Véronique;Jeffrey,StefanieS;Parker,PeterJ;Larijani,Banafshé
Akt/PKB 通路失调与多种人类癌症的不良预后相关。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较差的特异性的限制。在这里，我们报告了一种使用放大的时间分辨福斯特共振能量转移（FRET）的新型检测方法的开发，该检测方法具有高度特异性和敏感性，并且可以适应任何蛋白质。使用这种方法分析原代乳腺组织微阵列，我们以空间分辨率量化了激活的 pAkt 水平，揭示了肿瘤内的分子异质性。通过放大 FRET 评估的高 pAkt 状态与较差的无病生存率相关。我们的研究结果支持使用扩增的 FRET 来确定癌症组织中的 pAkt 状态，作为识别高危患者的候选生物标志物。 74(18)； 4983–95。 ©2014 AACR。
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.