Zhang Ya, Zeng Ruichao, Xia Yuanhang, Han Wei, Luan Yifei, Zhang Yuheng, Wu Shijia, Wang Shouhao, Wang Jinyong, Chen Yongping, Chen Dazhi
Hepatology Diagnosis and Treatment Center, The First Affiliated Hospital of Wenzhou Medical University & Zhejiang Provincial Key Laboratory for Accurate Diagnosis and Treatment of Chronic Liver Diseases, Wenzhou, 325035, China.
Hepatology Institute of Wenzhou Medical University, Wenzhou, 325035, China.
Mikrochim Acta. 2025 Mar 18;192(4):241. doi: 10.1007/s00604-025-06988-3.
Cancer cell metastasis is one of the major causes of patients death with hepatocellular carcinoma (HCC). Previous findings demonstrated that protrusion-accumulated STAT3 mRNA is highly related to HCC cell metastasis, making protrusion-localized STAT3 mRNA an ideal biomarker for evaluating HCC cell initiation and progression. A self-powered lipidic nanoflare (SLNF) has been developed for detecting the expression level of protrusion-accumulated STAT3 mRNA in individual HCC cells, which enables accurate prediction of HCC metastasis. The LNF system is a cholesterol micelle decorated with two kinds of DNA probes, a double-stranded response DNA and a single-stranded fuel probe. The cholesterol micelle can be easily assembled from an amphipathic cholesterol-conjugated DNA via hydrophobicity-mediated aggregation, exhibiting a highly efficient cell internalization. Moreover, the compact and high-density arrangement of DNA probes on the surface of cholesterol micelle enhances their biostability. All the above features make the LNF system an ideal approach for intracellular RNA imaging. The assay commences with the binding of STAT3 mRNA to the response DNA, which peels off the waste DNA and exposes the toehold domain. This domain serves as the proximal holding point for the fuel probe to initiate a strand displacement amplification, which is a crucial step in enabling the detection of targets expressed at trace levels, yielding a limit of detection (LOD) of 100 pM at 37 °C within 1.5 h. The SLNF system is expected to provide useful insight into the development of simple and degradation-resistant DNA probes for visual prediction of HCC metastasis, showing potential applications in tumor diagnosis and treatment.
癌细胞转移是肝细胞癌（HCC）患者死亡的主要原因之一。先前的研究结果表明，突出积累的STAT3 mRNA与HCC细胞转移高度相关，使得突出定位的STAT3 mRNA成为评估HCC细胞起始和进展的理想生物标志物。一种自供能脂质纳米耀斑（SLNF）已被开发用于检测单个HCC细胞中突出积累的STAT3 mRNA的表达水平，从而能够准确预测HCC转移。脂质纳米耀斑（LNF）系统是一种用两种DNA探针修饰的胆固醇胶束，一种是双链响应DNA，另一种是单链燃料探针。胆固醇胶束可以通过两亲性胆固醇共轭DNA经疏水性介导的聚集轻松组装而成，表现出高效的细胞内化。此外，DNA探针在胆固醇胶束表面的紧密且高密度排列增强了它们的生物稳定性。上述所有特性使LNF系统成为细胞内RNA成像的理想方法。该检测方法始于STAT3 mRNA与响应DNA的结合，这会剥离废弃DNA并暴露引发结构域。该结构域作为燃料探针启动链置换扩增的近端固定点，这是检测痕量水平表达的靶标的关键步骤，在37℃下1.5小时内检测限（LOD）为100 pM。SLNF系统有望为开发用于可视化预测HCC转移的简单且抗降解的DNA探针提供有用的见解，在肿瘤诊断和治疗中显示出潜在应用。
