Carbon Dot Based, Naphthalimide Coupled FRET Pair for Highly Selective Ratiometric Detection of Thioredoxin Reductase and Cancer Screening

Abstract

The fluorescence resonance energy transfer (FRET) mechanism has been established between carbon dots (CDs) and naphthalimide to monitor the activity of thioredoxin reductase (TrxR), which is often overexpressed in many cancer cells. The naphthalimide moiety was covalently attached to the surface of CDs through a disulfide linkage. In normal cell conditions (when devoid of high concentrations of TrxR), the CDs act as an energy donor and naphthalimide acts as an acceptor, which establishes the FRET pair as interpreted from the emission at λem = 565 nm, when excited at λex = 360 nm. However, contrary to this, the elevated levels of TrxR cause the breakage of disulfide bonds and consequently abolishes the FRET pair through the release of the naphthalimide moiety from the surface of CDs. This process was studied by monitoring of fluorescence intensity at λem = 565 and 440 nm, when excited at the same wavelength (λex = 360 nm). The TrxR based ratiometric quenching and enhancement of fluorescence intensity offers an interesting opportunity to monitor the enzyme activities and has many advantages over conventional monitoring of fluorescence intensity at a single wavelength to avoid interference of external factors. Fluorescence images of cancer cells in response to the nanosensor were visualized under a confocal microscope. Cytotoxicity study of nanosensor retards the growth of HeLa and MCF-7 cell lines in the presence of visible light. Therefore, the nanosensor also acts as a theranostic agent to diagnose as well as killing of cancer cells.