Institute for Functional Biomolecules, School of Chemistry and Chemical Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, 250353, PR China.
Textile Industrial Products Testing Center of Nanjing Customs District, Wuxi, 214101, PR China.
Anal Chim Acta. 2024 May 8;1302:342502. doi: 10.1016/j.aca.2024.342502. Epub 2024 Mar 19.
Cancer is a highly fatal disease which is close relative of miRNA aberrant expression and apoptosis disorders. Elucidation of the therapeutic efficacy through investigating the changes in miRNA and apoptosis holds immense importance in advancing the development of miRNA-based precision therapy. However, it remains a challenge as how to visually evaluate the efficacy during protocol optimization of miRNA-based anticancer drugs at the cellular level. Therefore, exploring effective and noninvasive methods for real-time monitoring of therapeutic efficacy in living cells is of great significance.
Herein, we reported a novel fluorescent nanoprobe COF-H1/H2-Peptide for visually evaluating drug efficacy in living cells through amplified imaging of low-abundant miRNA-221 with catalytic hairpin assembly (CHA) circle amplification, as well as simultaneous caspase-3 imaging. With strong stability and good biocompatibility, this newly fabricated amplified nanoprobe showed high sensitivity and specificity for the detection of miRNA-221 and caspase-3, and the limit of detection (LOD) of miRNA-221 was as low as 2.79 pM. The fluorescent imaging results showed that this amplified nanoprobe could not only detect caspase-3 in living cells, but also effectively detect low levels of miRNA-221 with increasing anticancer drug concentration and treatment time. The smart nanoprobe had effective performance for optimizing miRNA-based drug treatment schedules by dual-color fluorescence imaging.
This nanoprobe combined CHA amplified detection of intracellular miRNA-221 and synchronous apoptosis imaging, with excellent sensitivity for the detection of cellular low-level miRNA, enabling the realization of real-time assessment of the efficacy of miRNA-based therapy in living cells. This work presents a promising approach for revealing the regulatory mechanisms between miRNAs and apoptosis in cancer occurrence, development, and treatment.
癌症是一种高致命性疾病，与 miRNA 异常表达和细胞凋亡紊乱密切相关。阐明 miRNA 变化与细胞凋亡对治疗效果的影响，对于推进 miRNA 为基础的精准治疗发展具有重要意义。然而，如何在 miRNA 抗癌药物的方案优化过程中，在细胞水平上直观评估疗效，仍然是一个挑战。因此，探索有效的、非侵入性的方法，实时监测活细胞中的治疗效果具有重要意义。
本文报道了一种新型荧光纳米探针 COF-H1/H2-Peptide，通过催化发夹组装（CHA）循环扩增放大低丰度 miRNA-221 的成像，以及同时进行 caspase-3 成像，可用于直观评估活细胞中的药物疗效。该新构建的放大纳米探针具有强稳定性和良好的生物相容性，对 miRNA-221 和 caspase-3 的检测具有高灵敏度和特异性，miRNA-221 的检测限（LOD）低至 2.79 pM。荧光成像结果表明，该放大纳米探针不仅可以检测活细胞中的 caspase-3，而且可以随着抗癌药物浓度和治疗时间的增加，有效地检测低水平的 miRNA-221。该智能纳米探针通过双色荧光成像，对 miRNA 为基础的药物治疗方案的优化具有有效性能。
该纳米探针结合了细胞内 miRNA-221 的 CHA 放大检测和同步细胞凋亡成像，对细胞内低水平 miRNA 的检测具有优异的灵敏度，实现了实时评估 miRNA 为基础的治疗在活细胞中的疗效。这项工作为揭示 miRNA 与细胞凋亡在癌症发生、发展和治疗中的调控机制提供了一种有前景的方法。
