研究目的
Developing an effective method for detecting NQO1 activity with high sensitivity and selectivity in tumors for cancer diagnosis, treatment, and management.
研究成果
NQ-DCP is an effective fluorescent probe for non-invasive and real-time imaging of hNQO1 activity in cancer cells and tumors, with high selectivity, large Stokes shift, and good biocompatibility, showing promise for cancer bioimaging applications.
研究不足
The probe could not cross the blood-brain barrier for systemic imaging of intracranial tumors, requiring direct intracranial injection. In vivo applications may be limited by biodistribution and potential interferences in complex biological environments.
1：Experimental Design and Method Selection:
The study involved designing a fluorescent probe (NQ-DCP) based on dicyanoisophorone conjugated with quinone propionic acid, utilizing the 'quinone trimethyl lock system' for hNQO1 activation. Spectroscopic methods, HPLC analysis, fluorescence microscopy, flow cytometry, and in vivo imaging were employed.
2：Sample Selection and Data Sources:
Human cancer cell lines (DAOY, T98G, U87MG) and normal cells (astrocytes) were used, along with nude mice for xenograft models.
3：List of Experimental Equipment and Materials:
Instruments included Hitachi F-2500 Fluorescence spectrophotometer, Agilent HPLC instrument, IVIS Lumina XR Imaging system, BD LSRFortessa cell analyzer, Bio-rad ZOE fluorescent cell imager. Chemicals were from Sigma-Aldrich and Fisher Scientific.
4：Experimental Procedures and Operational Workflow:
Synthesis of NQ-DCP, spectroscopic measurements with hNQO1 and NADH, selectivity tests with various enzymes, cytotoxicity assays, cellular imaging, flow cytometry, in vivo injections and imaging in mice.
5：Data Analysis Methods:
Fluorescence intensity measurements, linear fitting for sensitivity, statistical analysis of mean fluorescence intensities, and image analysis using Living Image software.
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