One of the critical problems in bladder cancer (BC) management is the local recurrence of disease. However, achieving the accurate delineation of tumor margins intraoperatively remains extremely difficult due to the lack of effective tumor margin recognition technology. Herein, survivin molecular beacon (MB) and R11 peptide-linked spherical nucleic acids (SNAs) were synthesized as nanoprobes (AuNP-MB@R11) for sensitive detection of BC margins. Physicochemical properties proved that R11 peptides and survivin MB were successfully loaded onto the surface of SNAs. AuNP-MB@R11 had good stability against nuclease activity and high sensitivity and specificity to detect survivin single strand DNA (ssDNA) in vitro. According to cytology, R11 peptides could increase the BC targeting ability and membrane penetrability of SNAs. Notably, R11 peptides significantly promoted the disintegration of lysosomes and the release of SNAs to enhance fluorescence imaging quality. Further RNA sequencing proved that some genes and pathways related to endocytosis and lysosomes were significantly regulated, such as AGPAT5, GPD1L, and GRB2. In orthotopic BC models and a clinical sample from a patient with BC, AuNP-MB@R11 showed a more legible cancerous fluorescence margin and offered remarkably improved detection compared to those achieved by SNAs. R11 peptide-linked SNAs present promising potential to identify BC margin, which may help to improve the R0 resection rate in surgery and improve patients’ quality of life.

膀胱癌 (BC) 管理的关键问题之一是疾病的局部复发。然而，由于缺乏有效的肿瘤边缘识别技术，在术中准确勾画肿瘤边缘仍然极其困难。在本文中，存活蛋白分子信标（MB）和 R11 肽连接的球形核酸（SNA）被合成为纳米探针（AuNP-MB@R11），用于灵敏检测 BC 边缘。理化性质证明 R11 肽和存活蛋白 MB 成功加载到 SNA 表面。AuNP-MB@R11对核酸酶活性具有良好的稳定性，体外检测survivin单链DNA（ssDNA）的灵敏度和特异性高. 根据细胞学，R11肽可以增加SNA的BC靶向能力和膜渗透性。值得注意的是，R11 肽显着促进溶酶体的分解和 SNA 的释放，以提高荧光成像质量。进一步的RNA测序证明，一些与内吞作用和溶酶体相关的基因和通路受到显着调控，如AGPAT5、GPD1L和GRB2。在原位 BC 模型和来自 BC 患者的临床样本中，AuNP-MB@R11 显示出更清晰的癌性荧光边缘，并且与 SNA 实现的检测相比，提供了显着改善的检测。R11 肽连接 SNA 具有识别 BC 边缘的潜力，这可能有助于提高手术中的 R0 切除率并改善患者的生活质量。