Luo Xianzhu, Hu Erzhuo, Deng Fei, Zhang Cuiling, Xian Yuezhong
Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, Department of Chemistry, School of Chemistry and Molecular Engineering, East China Normal University Shanghai 200241 China
Key Laboratory of Haikou Trauma, Key Laboratory of Hainan Trauma and Disaster Rescue, Key Laboratory of Emergency and Trauma, Ministry of Education, The First Affiliated Hospital of Hainan Medical University, Hainan Medical University Haikou 571199 China.
Chem Sci. 2025 Mar 11;16(15):6507-6514. doi: 10.1039/d5sc00103j. eCollection 2025 Apr 9.
Precise recognition of senescent cells is essential owing to their key role in various diseases, including aging and tumor suppression. Although senescence-associated β-galactosidase (SA-β-Gal) is widely used as a senescence biomarker, it is not remarkably accurate due to its overexpression in some non-senescent cells. Herein, we developed a dual-channel fluorescent probe to improve the identification accuracy of senescent cells through simultaneous detection of β-gal and α-l-fucosidase (AFU) because the two markers are upregulated in senescent cells. The dual-channel fluorescent probe named HDQ-NA-AFU-Gal was employed to detect β-gal and AFU and identify senescence in living cells and tumor-bearing mice. When the two are present, the dual-enzyme activated probe emits strong red and green fluorescence at 740 nm and 550 nm, respectively, enabling independent detection of β-gal and AFU. This dual-enzyme detection approach allows for the precise differentiation between normal and senescent cells, particularly in ovarian cancer cells overexpressing β-gal. Furthermore, the probe can be applied as an effective tool for tracing β-gal and AFU during tumor senescence in mice. Thus, the dual-enzyme-responsive fluorescent probe has promising applications in biological research and clinical medicine.
由于衰老细胞在包括衰老和肿瘤抑制在内的各种疾病中起关键作用，因此对其进行精确识别至关重要。尽管衰老相关β-半乳糖苷酶（SA-β-Gal）被广泛用作衰老生物标志物，但由于其在一些非衰老细胞中的过度表达，其准确性并不显著。在此，我们开发了一种双通道荧光探针，通过同时检测β-半乳糖苷酶（β-gal）和α-L-岩藻糖苷酶（AFU）来提高衰老细胞的识别准确性，因为这两种标志物在衰老细胞中上调。名为HDQ-NA-AFU-Gal的双通道荧光探针用于检测β-gal和AFU，并识别活细胞和荷瘤小鼠中的衰老情况。当两者都存在时，双酶激活探针分别在740nm和550nm处发出强烈的红色和绿色荧光，从而能够独立检测β-gal和AFU。这种双酶检测方法能够精确区分正常细胞和衰老细胞，特别是在过表达β-gal的卵巢癌细胞中。此外，该探针可作为在小鼠肿瘤衰老过程中追踪β-gal和AFU的有效工具。因此，这种双酶响应荧光探针在生物学研究和临床医学中具有广阔的应用前景。
