NHC Key Laboratory of Birth Defects and Reproductive Health, Center of Smart Laboratory and Molecular Medicine, Fuling Hospital, School of Medicine, Chongqing University, Chongqing, 400044, PR China.
School of Clinical Medicine, Southwest Medical University, Luzhou, Sichuan, 646000, PR China.
Biosens Bioelectron. 2024 Oct 1;261:116496. doi: 10.1016/j.bios.2024.116496. Epub 2024 Jun 12.
Accurate intracellular visualization of human telomerase RNA (hTR) is imperative for early diagnosis and treatment monitoring of hepatocellular carcinoma (HCC). While isothermal amplification-based DNA cascade strategies are promising, challenges persist in achieving great intake efficiency of detection probes within tumor cells and enhancing intracellular reaction efficiency. This study introduces a SA@Comb-HCR nanosystem, a highly effective approach for in situ hTR detection in HCC cells. Sodium alginate-coated liposomes ensures efficient nanoprobe delivery, which are then combined with proximity effect-inspired signal amplification. The coating of sodium alginate facilitates receptor-mediated endocytosis, prevents serum protein adhesion, and mitigates cationic liposome cytotoxicity. The designed Comb-like consolidated hairpin probe enhances the concentration of the local reactant, resulting in cascade amplification upon hTR activation. This technique achieves precision detection of intracellularly overexpressed hTR in HCC cells with a remarkable detection limit of 0.7 pM. This approach holds great promise for advancing targeted and sensitive early clinical diagnosis of HCC.
准确的人端粒酶 RNA（hTR）细胞内可视化对于肝细胞癌（HCC）的早期诊断和治疗监测至关重要。虽然基于等温扩增的 DNA 级联策略很有前景，但在肿瘤细胞内实现高检测探针摄取效率和增强细胞内反应效率方面仍然存在挑战。本研究介绍了一种 SA@Comb-HCR 纳米系统，这是一种在 HCC 细胞中进行原位 hTR 检测的高效方法。海藻酸钠包被的脂质体确保了纳米探针的有效递送，然后与基于邻近效应的信号放大相结合。海藻酸钠的涂层促进了受体介导的内吞作用，防止了血清蛋白的黏附，并减轻了阳离子脂质体的细胞毒性。设计的 Comb 型整合发夹探针增强了局部反应物的浓度，在 hTR 激活时引发级联扩增。该技术实现了 HCC 细胞内过表达 hTR 的精确检测，检测限低至 0.7 pM。该方法有望推进 HCC 的靶向和敏感的早期临床诊断。
