Three-dimensional spheroids of non-malignant MCF10A and malignant SKBR3 breast cells were used for subsequent 3D Cell-SELEX to generate aptamers for specific binding and treatment of
breast cancer cells. Using 3D Cell-SELEX combined with Next-Generation Sequencing and bioinformatics, ten abundant aptamer families with specific structures were identified that selectively bind to SKBR3, and not to MCF10A cells. Multivalent aptamer
polymers were synthesized by co-polymerization and analyzed for binding performance as well as therapeutic efficacy. Binding performance was determined by confocal fluorescence imaging and revealed specific binding and efficient internalization of aptamer
polymers into SKBR3 spheroids. For therapeutic purposes, DNA sequences that intercalate the cytotoxic
drug doxorubicin were co-polymerized into the aptamer
polymers. Viability tests show that the
drug-loaded
polymers are specific and effective in killing SKBR3
breast cancer cells. Thus, the 3D-selected aptamers enhanced the specificity of
doxorubicin against malignant over non-malignant breast cells. The innovative modular
DNA aptamer platform based on 3D Cell SELEX and
polymer multivalency holds great promise for diagnostics and treatment of
breast cancer.