State Key Laboratory of Fine Chemicals, Dalian University of Technology, 2 Linggong Road, Dalian 116024, PR China.
School of Life Science and Biotechnology, Dalian University of Technology, 2 Linggong Road, Dalian 116024, PR China.
Biomaterials. 2018 Oct;179:1-14. doi: 10.1016/j.biomaterials.2018.06.028. Epub 2018 Jun 21.
Development of high selectivity, accurate targeted and noninvasive fluorescent probe for monitoring specific enzyme activity associated with the tumor is urgent needed for early diagnosis of cancer and clinical fluorescence interventional resection guidance treatment. Owing to the invasion of malignant tumor cells, tumor cells are mixed with normal cells in the actual tumor location, which make it quite difficult for clinician to diagnose early diagnosis of tumor as well as resection of tumor. To overcome aforementioned obstacle, herein, an ingenious enzyme-activated one and two-photon fluorescent probe TCF-GGT was constructed and synthesized including γ-GGT enzyme specific identification site and far-red fluorophore for imaging. Under simulative physiological condition, probe TCF-GGT demonstrated high selectivity, sensitivity (DL 0.014 mU/mL), rapid response (T 14 min) for the detection of γ-GGT enzyme. By virtue of its biocompatibility, probe was employed for the identification of ovarian cancer cells (A2780 cells) from normal cells (NIH-3T3 cells), particularly in mixed cultivation dish (simulate the actual environment of tumor) through 2D&3D fluorescence imaging with "dual" mark (Nucleic acid labeling used by Hoechst 33342 dye and γ-GGT enzyme labeling used by probe TCF-GGT) for the first time. Probe TCF-GGT could be visualize endogenous γ-GGT activity in HepG-2 cells and zebrafish on the two-photon confocal platform, which is conduce to estimate the inhibitor of γ-GGT enzyme in vivo. Through NaBu (a potential anticancer drug) stimulation, the changes of γ-GGT activity were observed in living MCF-7 cells by using this probe. More importantly, the deep tissue penetration ability of far-red fluorescence allowed the two-photon fluorescent probe TCF-GGT to real-time track γ-GGT activity in tissue slices and tumor xenotransplantation model of mice by the tail vein injection, which showed that this enzyme-triggered fluorogenic probe would be a potential tool for preclinical applications of tumor resection.
开发高选择性、准确靶向和非侵入性的荧光探针，用于监测与肿瘤相关的特定酶活性，对于癌症的早期诊断和临床荧光介入切除治疗至关重要。由于恶性肿瘤细胞的侵袭，肿瘤细胞与实际肿瘤部位的正常细胞混合在一起，这使得临床医生很难早期诊断肿瘤以及切除肿瘤。为了克服上述障碍，本文构建并合成了一种巧妙的酶激活型双光子荧光探针 TCF-GGT，其中包括γ-GGT 酶特异性识别位点和用于成像的远红荧光团。在模拟生理条件下，探针 TCF-GGT 对 γ-GGT 酶的检测表现出高选择性、灵敏度（DL 为 0.014 mU/mL）和快速响应（T 为 14 min）。由于其生物相容性，探针被用于从正常细胞（NIH-3T3 细胞）中识别卵巢癌细胞（A2780 细胞），特别是在混合培养盘中（模拟肿瘤的实际环境），通过二维和三维荧光成像进行“双重”标记（核酸标记使用 Hoechst 33342 染料，γ-GGT 酶标记使用探针 TCF-GGT）。探针 TCF-GGT 可以在双光子共聚焦平台上可视化 HepG-2 细胞和斑马鱼内源性 γ-GGT 活性，这有助于评估体内 γ-GGT 酶的抑制剂。通过 NaBu（一种潜在的抗癌药物）刺激，使用该探针可以观察到活 MCF-7 细胞中 γ-GGT 活性的变化。更重要的是，远红荧光的深组织穿透能力使得双光子荧光探针 TCF-GGT 能够通过尾静脉注射实时跟踪组织切片和小鼠肿瘤异种移植模型中的 γ-GGT 活性，这表明这种酶触发的荧光探针将成为肿瘤切除临床前应用的潜在工具。
