title: Oxidoreductase-Facilitated Visualization and Detection of Human Cancer Cells.
abstract: Achieving highly selective and sensitive detection/visualization of intracellular biological events through the use of cell-penetrable, bioanalyte-activatable, turn-on probes is dependent on the presence of specific event-linked cellular biomarkers, if and only if there exist activatable probes that appropriately respond to the biomarker analyte. Here is described the evaluation of, and use in cellular imaging studies, a previously undisclosed naphthalimide probe QMeNN, whose fluorescence is deactivated by photoinduced electron transfer (PeT) quenching that results from the presence of a covalently linked biomarker-specific quinone trigger group. Highly selective and rapid activation of the quinone group by the human cancer tumor-linked
  quinone oxido-reductase isozyme 1 (hNQO1) results in fast trigger group removal to yield a highly fluorescent green-energy-range reporter that possesses a high molar absorptivity; there is a 136-fold increase in brightness for the enzymatically produced reporter versus probe precursor, a value 4 times greater than previously reported for the hNQO1 analyte. The novel probe is taken up and activated rapidly within only hNQO1-positive human cancer cells; addition of an hNQO1 inhibitor prevents the selective activation of the probe. Comparison of cytosolic fluorescence intensity in positive cells versus background in negative cells yields a quantitative metric (positive-to-negative ratio, PNR) for judging hNQO1 activity. We show it is possible to determine hNQO1 presence in previously studied colorectal cancer cells and the unexplored ovarian cancer cell line NIH:OVCAR-3, with respective PNR values of 926 and 34 being obtained. Even with 10 min probe incubation, ready discrimination of positive cells from negative cells is achieved. Cell viability is unaffected by probe presence, thereby highlighting the practicality of probe use in live-cell imaging applications.
authors.affiliation: Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803-1804, United States.
authors.forename: Bijeta
authors.initials: B
authors.lastname: Prasai
authors.affiliation: Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803-1804, United States.
authors.forename: William C
authors.initials: WC
authors.lastname: Silvers
authors.affiliation: Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803-1804, United States.
authors.forename: Robin L
authors.initials: RL
authors.lastname: McCarley
mesh_terms: D002470:Cell Survival; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D009243:NAD / Q000378:metabolism; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000032:analysis / Q000378:metabolism; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology*; D010088:Oxidoreductases / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D005456:Fluorescent Dyes; D009243:NAD; D010088:Oxidoreductases; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human