1] Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan [2] Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Nat Commun. 2015 Mar 13;6:6463. doi: 10.1038/ncomms7463.
Fluorescence-guided diagnostics is one of the most promising approaches for facile detection of cancer in situ. Here we focus on β-galactosidase, which is overexpressed in primary ovarian cancers, as a molecular target for visualizing peritoneal metastases from ovarian cancers. As existing fluorescence probes are unsuitable, we have designed membrane-permeable HMRef-βGal, in which the optimized intramolecular spirocyclic function affords >1,400-fold fluorescence enhancement on activation. We confirm that HMRef-βGal sensitively detects intracellular β-galactosidase activity in several ovarian cancer lines. In vivo, this probe visualizes metastases as small as <1 mm in diameter in seven mouse models of disseminated human peritoneal ovarian cancer (SHIN3, SKOV3, OVK18, OVCAR3, OVCAR4, OVCAR5 and OVCAR8). Because of its high brightness, real-time detection of metastases with the naked eye is possible. Endoscopic fluorescence detection of metastases is also demonstrated. The results clearly indicate preclinical potential value of the probe for fluorescence-guided diagnosis of peritoneal metastases from ovarian cancers.
荧光引导诊断是简便原位检测癌症的最有前途的方法之一。在这里，我们专注于β-半乳糖苷酶，其在原发性卵巢癌中过度表达，作为可视化卵巢癌腹膜转移的分子靶标。由于现有的荧光探针不合适，我们设计了膜通透性 HMRef-βGal，其中优化的分子内螺环功能在激活时提供 >1400 倍的荧光增强。我们证实 HMRef-βGal 灵敏地检测几种卵巢癌细胞系中的细胞内β-半乳糖苷酶活性。在体内，该探针在七种传播性人腹膜卵巢癌（SHIN3、SKOV3、OVK18、OVCAR3、OVCAR4、OVCAR5 和 OVCAR8）的小鼠模型中可以检测到直径小至 <1mm 的转移灶。由于其高亮度，可通过肉眼实时检测转移灶。还证明了内窥镜荧光检测转移灶的能力。结果清楚地表明，该探针具有用于荧光引导诊断卵巢癌腹膜转移的临床前潜在价值。
