Miles James, Applebee Christopher J, Leboucher Pierre, Lopez-Fernandez Sonia, Lee Dae-Jin, Guarch Rosa, Ward Stephen, Parker Peter J, López Jose I, Larijani Banafshé
Cell Biophysics Laboratory, Ikerbasque, Basque Foundation for Science, FASTBASE SOLUTIONS Ltd, Research Centre for Experimental Marine Biology and Biotechnology (PiE) & Biofísika Institute (UPV/EHU, CSIC), University of the Basque Country, Spain.
Department of Pharmacy and Pharmacology, University of Bath, UK.
BBA Clin. 2017 Oct 13;8:97-102. doi: 10.1016/j.bbacli.2017.10.002. eCollection 2017 Dec.
Clear cell Renal Cell Carcinomas (ccRCC), the largest group of renal tumours, are resistant to classical therapies. The determination of the functional state of actionable biomarkers for the assessment of these adenocarcinomas is essential. The dysregulation of the oncoprotein, PKB/Akt has been linked with poor prognoses in human cancers.
MATERIAL & METHODS: We analysed the status of the PKB/Akt pathway in a representative tumour tissue microarray obtained from the primary tumours and their metastases in 60 ccRCC with long term follow up. We sought to define the evolution of this pathway from the primary tumour to the metastatic event and to know the impact of its functional state in tumour aggressiveness and patient survival. Two-site time resolved amplified FRET (A-FRET) was utilised for assessing the activation state of PKB/Akt and this was compared to conventional immunohistochemistry measurements.
Activation state of PKB/Akt in primary tumours defined by A-FRET correlated with poorer overall survival (hazard ratio 0.228; = 0.002). Whereas, increased protein expression of phosphoPKB/Akt, identified using classical immunohistochemistry, yielded no significant difference (hazard ratio 1.390; = 0.548).
Quantitative determination of PKB/Akt activation in ccRCC primary tumours alongside other diagnostics tools could prove key in taking oncologists closer to an efficient personalised therapy in ccRCC patients.
The quantitative imaging technology based on Amplified-FRET can rapidly analyse protein activation states and molecular interactions. It could be used for prognosis and assess drug function during the early cycles of chemotherapy. It enables evaluation of clinical efficiency of personalised cancer treatment.
透明细胞肾细胞癌（ccRCC）是最大的肾肿瘤群体，对传统疗法具有抗性。确定可用于评估这些腺癌的可操作生物标志物的功能状态至关重要。癌蛋白PKB/Akt的失调与人类癌症的不良预后相关。
我们分析了从60例ccRCC的原发性肿瘤及其转移灶中获取的代表性肿瘤组织微阵列中PKB/Akt信号通路的状态，并进行了长期随访。我们试图确定该信号通路从原发性肿瘤到转移事件的演变情况，并了解其功能状态对肿瘤侵袭性和患者生存的影响。采用两点时间分辨荧光共振能量转移（A-FRET）技术评估PKB/Akt的激活状态，并与传统免疫组织化学测量结果进行比较。
通过A-FRET确定的原发性肿瘤中PKB/Akt的激活状态与较差的总生存率相关（风险比0.228；P = 0.002）。而使用经典免疫组织化学鉴定的磷酸化PKB/Akt蛋白表达增加，未产生显著差异（风险比1.390；P = 0.548）。
在ccRCC原发性肿瘤中定量测定PKB/Akt激活状态以及其他诊断工具，可能是肿瘤学家更接近ccRCC患者有效个性化治疗的关键。
基于放大荧光共振能量转移的定量成像技术可以快速分析蛋白质激活状态和分子相互作用。它可用于预后评估，并在化疗早期周期评估药物功能。它能够评估个性化癌症治疗的临床疗效。
