We described the selection of a novel nucleic acid antibody-like prostate cancer (PCa) that specifically binds to the single-stranded DNA molecule from a 277-nt fragment that may have been partially paired and bound to the PCA3 RNA conformational structure.
PCA3-277 aptamer ligands were obtained, and the best binding molecule, named CG3, was synthesized for validation. Aiming to prove its diagnostic utility, we used an apta-qPCR assay with CG3-aptamer conjugated to magnetic beads to capture PCA3 transcripts, which were amplified 97-fold and 7-fold higher than conventional qPCR in blood and tissue, respectively. Histopathologic analysis of 161 prostate biopsies arranged in a TMA and marked with biotin-labeled CG3-aptamer showed moderate staining in both cytoplasm and nucleus of PCa samples; in contrast, benign prostatic hyperplasia (BPH) samples presented strong nuclear staining (78% of the cases). No staining was observed in stromal cells.
In addition, using an apta-qPCR, we demonstrated that CG3-aptamer specifically recognizes the conformational PCA3-277 molecule and at least three other transcript variants, indicating that long non-coding RNA (lncRNA) is processed after transcription. We suggest that CG3-aptamer may be a useful PCa diagnostic tool. In addition, this molecule may be used in drug design and drug delivery for PCa therapy.
Sci Rep. 2015 Jul 15;5:12090. doi: 10.1038/srep12090.
Marangoni K1, Neves AF2, Rocha RM3, Faria PR4, Alves PT5, Souza AG5, Fujimura PT5, Santos FA5, Araújo TG5, Ward LS6, Goulart LR7.
1 1] Laboratory of Cancer Molecular Genetics, Faculty of Medical Sciences, State University of Campinas, SP, Brazil [2] Laboratory of Nanobiotechnology, Institute of Genetics and Biochemistry, Federal University of Uberlândia, MG, Brazil.
2 Laboratory of Molecular Biology, Institute of Biotechnology, Federal University of Goiás, Catalão/GO, Brazil.
3 AC Camargo Cancer Hospital, SP, Brazil.
4 Laboratory of Histology, Institute of Biomedical Sciences, Federal University of Uberlândia, MG, Brazil.
5 Laboratory of Nanobiotechnology, Institute of Genetics and Biochemistry, Federal University of Uberlândia, MG, Brazil.
6 Laboratory of Cancer Molecular Genetics, Faculty of Medical Sciences, State University of Campinas, SP, Brazil.
7 1] Laboratory of Nanobiotechnology, Institute of Genetics and Biochemistry, Federal University of Uberlândia, MG, Brazil [2] University of California-Davis, Dept. of Medical Microbiology and Immunology, Davis/CA, USA.