Department of Chemistry & State Key Laboratory of Molecular Engineering of Polymers & Institute of Biomedicine Science, Fudan University, Shanghai 200433, China.
Department of Obstetrics and Gynecology of Shanghai Medical School & Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases & Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai 200011, China.
Theranostics. 2019 Jun 24;9(16):4597-4607. doi: 10.7150/thno.35322. eCollection 2019.
During a minimally invasive tumor resection procedure, it is still a challenge to rapidly and accurately trace tiny malignant tumors in real time. Fluorescent molecular imaging is considered an efficient method of localizing tumors during surgery due to its high sensitivity and biosafety. On the basis of the fact that γ-glutamyltranspeptidase (GGT) is overexpressed in ovarian cancer, we herein designed a highly sensitive ratiometric fluorescent GGT-responsive probe Py-GSH for rapid tumor detection. The GGT response probe (Py-GSH) was constructed by using GSH group as a response group and pyrionin B as a fluorescent reporter. Py-GSH was characterized for photophysical properties, response speed and selectivity of GGT and response mechanism. The anti-interference ability of ratiometric probe Py-GSH to probe concentration and excitation power was evaluated both and in tissue. The biocompatibility and toxicity of the ratiometric probe was examined using cytoxicity test. The GGT levels in different lines of cells were determined by ratiometric fluorescence imaging and cytometry analysis. The obtained probe capable to rapidly monitored GGT activity in aqueous solution with 170-fold ratio change. By ratiometric fluorescence imaging, the probe Py-GSH was also successfully used to detect high GGT activity in solid tumor tissues and small peritoneal metastatic tumors (~1 mm in diameter) in a mouse model. In particular, this probe was further used to determine whether the tissue margin following clinical ovarian cancer surgery contained tumor. In combination of ratiometric fluorescence probes with imaging instrument, a point-of-care imaging method was developed and may be used for surgical navigation and rapid diagnosis of tumor tissue during clinical tumor resection.
在微创肿瘤切除手术中，实时快速准确地追踪微小恶性肿瘤仍然是一个挑战。荧光分子成像是一种有效的手术中定位肿瘤的方法，因为它具有高灵敏度和生物安全性。基于 γ-谷氨酰转肽酶（GGT）在卵巢癌中过度表达的事实，我们设计了一种高度敏感的比率荧光 GGT 响应探针 Py-GSH 用于快速肿瘤检测。GGT 响应探针（Py-GSH）由 GSH 基团作为响应基团和吡咯并因 B 作为荧光报告基团构建而成。对 GGT 和响应机制的光物理性质、响应速度和选择性进行了研究。评估了比率探针 Py-GSH 在组织内和组织内对探针浓度和激发功率的抗干扰能力。通过细胞毒性试验研究了比率探针的生物相容性和毒性。通过比率荧光成像和细胞术分析测定了不同细胞系中的 GGT 水平。该探针能够在水溶液中快速监测 GGT 活性，比率变化达 170 倍。通过比率荧光成像，探针 Py-GSH 还成功用于检测小鼠模型中固体肿瘤组织和小腹膜转移性肿瘤（直径约 1 毫米）中的高 GGT 活性。特别是，该探针进一步用于确定临床卵巢癌手术后的组织边缘是否含有肿瘤。结合比率荧光探针和成像仪器，开发了一种即时成像方法，可用于临床肿瘤切除过程中的手术导航和肿瘤组织的快速诊断。
