Evaluating internalization and recycling of folate receptors in breast cancer cells using quantum dots

Folic acid (FA) regulates metabolic activities essential to the human body. FA receptor (FR) overexpression has been reported for many cancers, but there are still few or conflicting data about FRs in breast cancer cells. Quantum dots (QDs) have arisen as tools to elucidate aspects on FRs, due to their unique physicochemical properties. Herein, QDs conjugated to FA were explored to study the internalization and recycling of FRs in breast cancer cells, using HeLa as an out-group control. QDs were covalently conjugated to FA under different conditions. The best conjugate was applied to study FRs in HeLa, MCF7, MDA-MB231, and T47D cells applying confocal microscopy and flow cytometry analyses. The conjugation efficiency and specificity were evaluated, respectively, using fluorescence correlation spectroscopy (FCS) and saturation assays. FCS confirmed the effectiveness of the conjugation. HeLa and T47D had/internalized a higher amount of FRs (95% and 90% of labeling, respectively) than MDA-MB231 cells (68%). MCF7 cells seem to have very low functional FRs (3%). Saturation assays proved the specificity of QD-FA conjugates and suggested that FR recycling rate is low in the majority of cells studied, except for T47D. QD-FA conjugates were successfully developed. Therapies targeting FRs may be more effective for HeLa, T47D, and MDA-MB231.