Construction Of A Glutathione Fluorescent Probe And Its Application In The Chemotherapy Evaluation Of Nasopharyngeal Carcinoma

Objective:Nasopharyngeal carcinoma(NPC)is a malignant tumor of the head and neck originating in the nasopharynx,commonly treated with radiation and chemotherapy.Clinically,cisplatin(DDP)-based chemotherapy regimens occupy an important position in the treatment of NPC,but DDP resistance affects the treatment effect of nasopharyngeal carcinoma.DDP resistance is closely related to glutathione(GSH)concentrations,mainly by affecting cisplatin transport and metabolism.Fluorescent probes have the advantages of real-time non-invasive visual imaging,and have strong advantages in disease diagnosis and efficacy monitoring.Therefore,in this study,we intend to design a fluorescent probe for the detection of intracellular GSH for real-time visualization and monitoring of the chemotherapy effect of NPC.Methods: The probe SNAFL-GSH is synthesized by chemical organic reaction,and the structure of the probe is characterized by nuclear magnetic hydrogen spectroscopy,nuclear magnetic carbon spectroscopy,and high-resolution mass spectrometry.Evaluate the spectral performance of the probe with a fluorescence spectrophotometer.The CCK-8 kit was used to evaluate the biosafety of probe.The sensitivity and specificity of SNAFL-GSH for the detection of GSH in vivo by adding N-ethylmaleimide and glutathione ethyl ester to the nasopharyngeal cell line was changed to change the GSH concentration.The potential of SNAFL-GSH to visualize the effect of NPC chemotherapy by adding DDP and Erastin,respectively,to detect cell viability and assess changes in intracellular GSH concentration with probes was added to nasopharyngeal cancer cell lines.By establishing a mouse model of nasopharyngeal carcinoma,the imaging ability of the probe in mice was understood.GSH in fresh frozen sections of clinical nasopharyngeal carcinoma was detected by probes,and the potential of probes to image GSH in clinical samples was explored.Results: The results of nuclear magnetic hydrogen spectroscopy,carbon spectroscopy and mass spectrometry showed that the synthesized probe was consistent with the designed structure.The spectral test results of SNAFL-GSH showed that SNAFL-GSH had good selectivity for GSH,and there was an excellent linear relationship between probe fluorescence signal change and GSH concentration change.Cytotoxicity experiments showed that the probe SNAFL-GSH had low toxicity and was suitable for biological experiments.The probe SNAFL-GSH imaged GSH in different nasopharyngeal cell lines,which proved that the GSH level in nasopharyngeal cancer cell lines was significantly higher than that of normal cells.The probe SNAFL-GSH can dynamically monitor the changes of intracellular GSH levels when different drugs treat nasopharyngeal cancer cell lines;Mouse nasopharyngeal carcinoma model experiments and clinical nasopharyngeal carcinoma tissue imaging experiments proved that the probe has excellent in vivo biological imaging performance.Conclusions: In this study,the fluorescent probe SNAFL-GSH was designed and synthesized,and SNAFL-GSH was confirmed by in vitro experiments that SNAFL-GSH has high selectivity and sensitivity to GSH.The results of SNAFL-GSH imaging in different live cell lines,mouse nasopharyngeal carcinoma models and human nasopharyngeal carcinoma tissues showed that SNAFL-GSH can be used as a powerful chemical tool to monitor the efficacy of chemotherapy for nasopharyngeal carcinoma.