研究目的
To evaluate a fluorescent-labelled erlotinib based theranostic agent for the molecular imaging of mutated EGFR tumours in vitro and ex vivo using a mice xenograft model and fibred confocal fluorescence microscopy (FCFM).
研究成果
The fluorescent erlotinib-based tracer can identify EGFR-mutated tumors with high sensitivity and specificity in real-time using FCFM, but further studies on human samples are needed for clinical application.
研究不足
The study did not perform standard confocal microscopy to confirm intracellular localization of the tracer, did not assess functional consequences of fluorescein addition, and could not distinguish sensitive from resistant EGFR mutations ex vivo. It was not tested on human samples, so cut-off values and background noise may differ.
1：Experimental Design and Method Selection:
The study involved synthesizing a fluorescent tracer by adding fluorescein to erlotinib, using cell lines with different EGFR statuses, and employing FCFM for real-time imaging. MTT assay was used to assess cell viability.
2：Sample Selection and Data Sources:
Four human adenocarcinoma cell lines (HCC827, H1975, H1650 with mutated EGFR, and A549 with wild-type EGFR) were obtained from ATTC and xenografted onto 35 Nude mice.
3：List of Experimental Equipment and Materials:
Equipment included a Cellvizio lab? system with optical probe, MTT assay kit, culture media (RPMI and DMEM), and chemicals for tracer synthesis. Materials included cell lines, mice, and reagents like PBS and fluorescein.
4：Experimental Procedures and Operational Workflow:
Cells were cultured, xenografted, and exposed to tracer. In vitro imaging on cell pellets and ex vivo imaging on excised tumors after immersion in tracer solution were performed. FCFM imaging was done with probe calibration and MFI recording.
5：Data Analysis Methods:
Statistical analysis used Student T-test, ANOVA, Spearman correlation, and ROC curves with GraphPad Prism software.
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