Visual screening and efficacy evaluation of high-performance fluorescent probe DAF-FM in esophagitis cancer transformation
Year: 2025
Type: article
Abstract: BACKGROUND Chronic esophagitis can progress to esophageal cancer via "inflammation-dysplasia-cancer" transformation, with nitric oxide (NO) serving as a critical mediator in this process. Traditional diagnostic methods (e.g. , endoscopic biopsy) for esophageal cancer transformation have low sensitivity and require long detection time, while existing fluorescent probes lack specificity and stability for real-time NO monitoring. High-performance fluorescent probes like DAF-FM, with NO-targeting ability, show potential for visual screening and efficacy evaluation but need systematic validation in esophageal cancer models. AIM To validate the applicability of the fluorescent probe DAF-FM for visual screening of esophageal cancer transformation, explore the underlying mechanism of NO-regulated transformation, and evaluate the probe’s efficacy in monitoring therapeutic responses. METHODS Laser confocal imaging and flow cytometry were used to analyze DAF-FM’s NO concentration/time-dependent fluorescence response, lysosomal targeting (via Pearson coefficient), and cytotoxicity (with cholecystokinin-8 assay) in esophageal cells. Sprague-Dawley rat esophageal cancer models (normal, esophagitis, esophageal cancer, and drug/radiotherapy intervention) were established to monitor NO dynamics and tumor volume correlation. Clinical diagnostic comparison (50 suspected patients) with endoscopic biopsy/histopathology was conducted using Kolmogorov-Smirnov test and Student’s t -test (P < 0.05). Western blot and quantitative real-time polymerase chain reaction were used to explore NO’s role in the nuclear factor-kappa B (NF-κB) pathway. RESULTS DAF-FM exhibited concentration/time-dependent fluorescence with NO (300 μM NO: 60-minute fluorescence intensity 458 ± 15 arbitrary units, P < 0.05) and specific lysosomal targeting (Pearson’s coefficient = 0.82 ± 0.03). It had low cytotoxicity (82.3% ± 4.1% cell viability at 50 μM). In rat models, DAF-FM showed that NO was correlated with tumor volume (R ² = 0.87). Clinically, its sensitivity (92.5%) outperformed endoscopic biopsy (78.3%), with shorter detection time (30 minutes vs 48 hours, P < 0.05). Mechanistically, NO regulated transformation via the NF-κB pathway (Pearson’s coefficient = 0.78 ± 0.05 between DAF-FM and NF-κB). CONCLUSION DAF-FM is a feasible tool for visual screening of esophageal cancer transformation, enabling real-time NO monitoring, high-sensitivity diagnosis, and therapeutic efficacy evaluation. It provides a new approach for esophageal cancer diagnosis and mechanism research.
Authors: Wan-Hua Chen, Cheng-Feng Cai, Bing-Zhong Gao, Wen-Shan Hong, Ying-Zhi Xu, Wen-Jie Cai
Institutions: Fujian Medical University, Ningxia Medical University
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: diamond
OpenAlex ID: W7117240651