GSH Activated Biotin-tagged Near-Infrared Probe for Efficient Cancer Imaging
GSH Activated Biotin-tagged Near-Infrared Probe for Efficient Cancer Imaging
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GSH 激活生物素标记近红外探针可实现高效癌症成像
DOI：
10.7150/thno.32742
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发表时间：
2019-01-01
期刊：
影响因子：
12.4
通讯作者：
Lin, Nengming
中科院分区：
文献类型：
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作者：
Guo, Ruiying;Huang, Feng;Lin, Nengming
在实体瘤的诊断和手术切除中，需要高特异性和高灵敏度的肿瘤成像工具来辅助边界识别。在这项研究中，我们开发了一种近红外（NIR）探针（P6）的基础上，在体外/体内肿瘤成像的双重战略的癌细胞靶向和刺激依赖性激活。研究了P6对癌细胞的选择性成像能力，并初步探讨了P6内吞的可能机制。方法：设计、合成并表征了GSH活化的生物素标记近红外探针P6。通过紫外-可见光谱、荧光光谱和LC-MS分析，系统地阐明了GSH的响应特性。在共聚焦激光扫描显微镜下，在各种活癌细胞系（即SW 480、HGC-27、H460、BxPC-3、KHOS）和正常细胞系（即BEAS-2B、HLF-1、THP 1）中收集探针P6的体外荧光成像。进一步应用探针P6对从腹膜癌和直肠癌患者新鲜分离的原代人癌细胞进行成像。收集人肿瘤组织的连续切片并送去进行H&E（苏木精-伊红）染色和P6成像。采用活体荧光和光声成像技术研究P6在HGC-27和KHOS异种移植瘤模型中肿瘤和正常组织中的体内成像。结果如下：探针P6可被肿瘤特异性生物素受体识别并转运至癌细胞内，并被GSH有效触发释放荧光团4。事实上，P6的细胞摄取可以通过加入游离生物素而部分阻断。此外，探针P6可以对各种癌细胞系以及原发性癌细胞成像，表现出比正常细胞高10倍的荧光强度。在新鲜解剖的癌组织中，P6荧光成像在共聚焦激光扫描显微镜下区分癌区域，其与H&E染色所指示的区域完全相同。我们还发现P6通过局部注射对癌组织表现出优越的上级选择性。结论：在这项研究中，我们开发了一种双模式近红外探针P6，具有增强的细胞摄取到癌细胞和环境刺激触发的荧光。我们的策略为成像工具的开发提供了新的见解，这些成像工具可能用于荧光图像引导的癌症边界识别和可能的癌症诊断。
Tumor imaging tools with high specificity and sensitivity are needed to aid the boundary recognition in solid tumor diagnosis and surgical resection. In this study, we developed a near infra-red (NIR) probe (P6) for in vitro/in vivo tumor imaging on the basis of the dual strategy of cancer cell targeting and stimulus-dependent activation. The selective imaging capacity towards cancer cells of P6 was thoroughly investigated, and the potential mechanisms of endocytosis were preliminary explored. Methods: GSH-activated biotin labelled NIR probe (P6) was designed, synthesized and characterized. The GSH responsive properties were systematically illustrated through UV-vis, fluorescent tests and LC-MS analysis. In vitro fluorescent imaging of probe P6 was collected in various living cancer cell lines (i.e. SW480, HGC-27, H460, BxPC-3, KHOS) and normal cell lines (i.e. BEAS-2B, HLF-1, THP1) under confocal laser scanning microscopy. Probe P6 was further applied to image primary human cancer cells which were freshly isolated from the peritoneal carcinoma and rectal cancer patients. Serial sections of human tumor tissues were collected and sent for H&E (hematoxylin-eosin) staining and P6 imaging. Live fluorescent and photoacoustic imaging were used to investigate the in vivo imaging of P6 in both tumor and normal tissues in HGC-27 and KHOS xenograft model. Results: Probe P6 could be recognized and transported into cancer cells by tumor specific biotin receptors and efficiently be triggered by GSH to release fluorophore 4. In fact, the cellular uptake of P6 could be partially blocked by the addition of free biotin. Furthermore, probe P6 could image various cancer cell lines, as well as primary cancer cells, exhibiting a ten-fold increase in fluorescence intensity over normal cells. In freshly dissected cancer tissues, P6 fluorescent imaging distinguished the cancerous area under confocal laser scanning microscopy, which was exact the same area as indicated by H&E staining. We also found that P6 exhibited superior selectivity against cancer tissues by local injection. Conclusion: In this study, we developed a dual-modal NIR probe P6 with enhanced cellular uptake into cancer cells and environmental stimulus triggered fluorescence. Our strategy provided a novel insight into the development of imaging tools that could be potentially used for fluorescent image-guided cancer boundary recognition and possibly cancer diagnosis.