In situ imaging of quantum dot-AZD4547 conjugates for tracking the dynamic behavior of fibroblast growth factor receptor 3

Abstract

Fibroblast growth factor receptors (FGFRs) play an important role in determining cell proliferation, differentiation, migration, and survival. Although a variety of small-molecule FGFR inhibitors have been developed for cancer therapeutics, the interaction between FGFRs and FGFR inhibitors has not been well characterized. The FGFR–inhibitor interaction can be characterized using a new imaging probe that has strong, stable signal properties for in situ cellular imaging of the interaction without quenching. We developed a kinase–inhibitor-modified quantum dot (QD) probe to investigate the interaction between FGFR and potential inhibitors. Especially, turbo-green fluorescent protein-FGFR3s were overexpressed in HeLa cells to investigate the colocalization of FGFR3 and AZD4547 using the QD-AZD4547 probe. The result indicates that this probe is useful for investigating the binding behaviors of FGFR3 with the FGFR inhibitor. Thus, this new inhibitor-modified QD probe is a promising tool for understanding the interaction between FGFR and inhibitors and for creating future high-content, cell-based drug screening strategies.

Tumor diagnosis using carbon-based quantum dots: Detection based on the hallmarks of cancer

“…FGFs are secreted glycoproteins that are sequestered in the ECM. Hwang et al [ 379 ] designed a kinase-inhibitor-modified QD probe to study the reaction between FGFRs and potential inhibitors. More specifically, turbo-green fluorescent protein-FGFR3s were overexpressed in HeLa cells to explore the colocalization of FGFR3 and AZD4547 using the QD probe.…”

Dissection of Protein Kinase Pathways in Live Cells Using Photoluminescent Probes: Surveillance or Interrogation?

“…Such approach suffers remarkably from the background signal of the unbound probe, which is only somewhat reduced by the repeated washing procedures. In some cases, fixation of cells was performed after application of probes to reduce the background signal and to confirm selectivity of probe binding by co-localization with immunofluorescent staining [177][178][179][180][181]. However, fixation is associated with disruption of membrane structures, which can give rise to significant artifacts in imaging: the localization of the probe before and after fixation may differ drastically.…”