使用纳米荧光素酶和萤火虫荧光素酶进行双报告基因成像检测脑肿瘤和全身转移灶
Detection of Brain Tumors and Systemic Metastases Using NanoLuc and Fluc for Dual Reporter Imaging.
作者信息
Germain-Genevois Coralie, Garandeau Olivia, Couillaud Franck
机构信息
Institut de Bio-Imagerie (IBIO), CNRS/UMS 3428, University Bordeaux, 146 rue Léo Saignat, 33076, Bordeaux cedex, France.
Centre de Résonance Magnétique des Systèmes Biologiques (RMSB), CNRS/UMR 5536, University Bordeaux, 146 rue Léo Saignat, 33076, Bordeaux cedex, France.
出版信息
Mol Imaging Biol. 2016 Feb;18(1):62-9. doi: 10.1007/s11307-015-0864-2.
PURPOSE
Bioluminescence imaging (BLI) is a technique with a low background noise and high sensitivity which is widely used in mice models in oncology. We aimed to assess BLI efficiency of the new luciferase NanoLuc (Nluc) for glioblastoma cell lines and tumors, including for dual reporter applications of deep brain tumors and systemic metastasis when combined with firefly luciferase (Fluc).
PROCEDURES
U87 cells were genetically modified for constitutive production of either Nluc, Fluc, or both and assayed for luciferase activity and BLI on cell lysates, living cells, subcutaneous tumors, brain tumors, and systemic metastases.
RESULTS
In vitro, light production by Nluc activity is higher than Fluc. In vivo, Nluc allows for tumor detection including for deep brain tumors and systemic metastases.
CONCLUSIONS
Nluc appears to be a useful tool to combine with Fluc for dual imaging in vivo using bioluminescence, allowing for the detection of distinct events in deep tissues within the same organism.
目的
生物发光成像（BLI）是一种背景噪声低、灵敏度高的技术，广泛应用于肿瘤学的小鼠模型中。我们旨在评估新型荧光素酶纳米荧光素酶（Nluc）对胶质母细胞瘤细胞系和肿瘤的BLI效率，包括与萤火虫荧光素酶（Fluc）联合用于深部脑肿瘤和全身转移的双报告基因应用时的效率。
程序
对U87细胞进行基因改造，使其组成性产生Nluc、Fluc或两者，并对细胞裂解物、活细胞、皮下肿瘤、脑肿瘤和全身转移灶进行荧光素酶活性和BLI检测。
结果
在体外，Nluc活性产生的光比Fluc高。在体内，Nluc可用于肿瘤检测，包括深部脑肿瘤和全身转移灶。
结论
Nluc似乎是一种与Fluc结合用于体内生物发光双成像的有用工具，可在同一生物体的深部组织中检测不同事件。
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