TYR as a multifunctional reporter gene regulated by the Tet-on system for multimodality imaging: an in vitro study.
TYR as a multifunctional reporter gene regulated by the Tet-on system for multimodality imaging: an in vitro study.
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TYR 作为受 Tet-on 系统调节的多功能报告基因，用于多模态成像：一项体外研究。
DOI：
10.1038/srep15502
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发表时间：
2015-10-20
影响因子：
4.6
通讯作者：
Lan X
中科院分区：
文献类型：
--
作者：
Feng H;Xia X;Li C;Song Y;Qin C;Zhang Y;Lan X
人酪氨酸酶基因TYR是一个多功能报告基因，在光声成像（派）、正电子发射断层扫描（PET）和磁共振成像（MRI）中具有潜在的用途。我们试图建立和评估一个报告基因系统，使用TYR的Tet-on基因表达系统（基因表达诱导的强力霉素[Dox]）的控制下，作为一种多模态成像剂。我们将TYR转染到人乳腺癌细胞（MDA-MB-231）中，将所得细胞系命名为231-TYR。使用未转染的MDA-MB-231细胞作为对照，我们使用蛋白质印迹、细胞酪氨酸酶活性、Masson-Fontana银染色和细胞免疫荧光研究验证了与Dox孵育后231-TYR成功表达TYR，而对照细胞和未暴露于Dox的231-TYR细胞显示没有TYR表达。通过在405 nm处的吸光度检测，黑色素浓度的增加与Dox浓度和孵育时间呈正相关。TYR表达的Dox诱导的转染细胞缩短MRI T1和T2弛豫时间。在这些细胞中很容易检测到光声信号。靶向黑色素的18F-5-氟-N-（2-[二乙基氨基]乙基）吡啶酰胺（18F-5-FPN）在Dox诱导的231-TYR细胞中迅速积累。这些表明TYR诱导黑色素产生受Tet-on系统调节，并且含有TYR的指示细胞可能在多模态成像中具有实用性。
The human tyrosinase gene TYR is a multifunctional reporter gene with potential use in photoacoustic imaging (PAI), positron emission tomography (PET), and magnetic resonance imaging (MRI). We sought to establish and evaluate a reporter gene system using TYR under the control of the Tet-on gene expression system (gene expression induced by doxycycline [Dox]) as a multimodality imaging agent. We transfected TYR into human breast cancer cells (MDA-MB-231), naming the resulting cell line 231-TYR. Using non-transfected MDA-MB-231 cells as a control, we verified successful expression of TYR by 231-TYR after incubation with Dox using western blot, cellular tyrosinase activity, Masson-Fontana silver staining, and a cell immunofluorescence study, while the control cells and 231-TYR cells without Dox exposure revealed no TYR expression. Detected by its absorbance at 405 nm, increasing concentrations of melanin correlated positively with Dox concentration and incubation time. TYR expression by Dox-induced transfected cells shortened MRI T1 and T2 relaxation times. Photoacoustic signals were easily detected in these cells. 18F-5-fluoro-N-(2-[diethylamino]ethyl)picolinamide (18F-5-FPN), which targets melanin, quickly accumulated in Dox-induced 231-TYR cells. These show that TYR induction of melanin production is regulated by the Tet-on system, and TYR-containing indicator cells may have utility in multimodality imaging.