Emerging trends on the uptake of fluorescent probes based on glucose analogs by cancer cells: From basic studies to therapeutics.

The cancer cell metabolism, notably characterized by the Warburg effect, has been the focus of intense investigation regarding the mechanisms of the uptake of glucose analogs, opening up perspectives for diagnosis and treatment of cancer disease. In this review, we delve into the ever-evolving landscape of cancer research, centering on fluorescent probes based on glucose analogs. These analogs, resulting from modifications in the carbohydrate structure with functional groups, have stood out as versatile molecules in applications ranging from disease comprehension to therapeutic innovation, especially when combined with fluorescent compounds. Fluorescence-based assays have provided valuable contributions to the revelation of complex biological mechanisms in life sciences. This review presents selected studies from about the past six years up to 2024 related to the use of glucose-based fluorescent probes, for the investigation of their uptake profile as well as for therapeutic purposes. We believe that these investigations offer insights into the intricate interaction between glucose analogs and cancer cell metabolism, guiding future research and clinical applications in this field.

A fluorescent glyconanoprobe based on quantum dots and thiolated glucose: Applications in monolayers and spheroids of cancer cells.

The differential energy metabolism of cancer cells has stimulated the development of tools that can be applied to better understand the complex biological interaction involved in the uptake of glucose analogs at the cellular level in this disease. Herein, we explored the outstanding optical properties of quantum dots (QDs) to develop a new fluorescent glyconanoprobe using the 1-thio-ß-d-glucose (Glc). Then, monolayers and spheroids of HeLa cells were applied to probe the biological interaction with the conjugate through fluorescence techniques. Spheroids have been gaining prominence for better mimicking the tumor microenvironment. The Glc-QDs conjugate was prepared by a facile and direct procedure based on the affinity of the Glc thiol group by the QD semiconductor surface. The conjugation was evaluated and confirmed by Zeta potential (ζ) measurements, FTIR spectroscopy, and fluorescence correlation spectroscopy (FCS). Moreover, a biological assay using Candida albicans yeasts coated with concanavalin A, by exploring the lectin-carbohydrate affinity, was also developed to further confirm the conjugation, which corroborated the previous analyses. The hanging drop method was used to prepare the spheroids. The fluorescence microscopy analyses indicated an intracellular labeling by the glyconanoprobe, in both cell culture models. Flow cytometry assays revealed effective uptake of the conjugate (above ca. 76%), even by cells cultivated as spheroids, applying short incubation time. Therefore, a new fluorescent glyconanoprobe was developed, which showed potential to be applied for investigating mechanisms involved in the uptake of glucose analogs, both by simpler and complex cancer biological models, as monolayers and spheroids.

Advances in the study of spheroids as versatile models to evaluate biological interactions of inorganic nanoparticles.

Spheroids are in vitro three-dimensional multicellular microstructures able to mimic the biological microenvironment, including the complexity of tumor architecture. Therefore, results closer to those expected for in vivo organisms can be reached using spheroids compared to the cell culture monolayer model. Inorganic nanoparticles (NPs) have also been playing relevant roles in the comprehension of biological processes. Moreover, they have been probed as novel diagnostic and therapeutical nanosystems. In this context, in this review, we present applications, published in the last five years, which show that spheroids can be versatile models to study and evaluate biological interactions involving inorganic NPs. Applications of spheroids associated with (i) basic studies to assess the penetration profile of nanostructures, (ii) the evaluation of NP toxicity, and (iii) NP-based therapeutical approaches are described. Fundamentals of spheroids and their formation methods are also included. We hope that this review can be a reference and guide future investigations related to this interesting three-dimensional biological model, favoring advances to Nanobiotechnology.