title: Screening and characterization of a novel RNA aptamer that specifically binds to human prostatic acid phosphatase and human prostate cancer cells.
abstract: Prostatic acid phosphatase (PAP) expression increases proportionally with prostate cancer progression, making it useful in prognosticating intermediate to high-risk prostate cancers. A novel ligand that can specifically bind to PAP would be very helpful for guiding prostate cancer therapy. RNA aptamers bind to target molecules with high specificity and have key advantages such as low immunogenicity and easy synthesis. Here, human PAP-specific aptamers were screened from a 2'-fluoropyrimidine (FY)-modified RNA library by SELEX. The candidate aptamer families were identified within six rounds followed by analysis of their sequences and PAP-specific binding. A gel shift assay was used to identify PAP binding aptamers and the 6N aptamer specifically bound to PAP with a Kd value of 118 nM. RT-PCR and fluorescence labeling analyses revealed that the 6N aptamer bound to PAP-positive mammalian cells, such as PC-3 and LNCaP. IMR-90 negative control cells did not bind the 6N aptamer. Systematic minimization analyses revealed that 50 nucleotide sequences and their two hairpin structures in the 6N 2'-FY RNA aptamer were equally important for PAP binding. Renewed interest in PAP combined with the versatility of RNA aptamers, including conjugation of anti-cancer drugs and nano-imaging probes, could open up a new route for early theragnosis of prostate cancer.
authors.affiliation: Department of Molecular Biology, Dankook University, Yongin 448-701, Korea ; Institute of Nanosensor and Biotechnology, Dankook University, Yongin 448-701, Korea.
authors.forename: Hoon Young
authors.initials: HY
authors.lastname: Kong
authors.affiliation: Department of Molecular Biology, Dankook University, Yongin 448-701, Korea ; Institute of Nanosensor and Biotechnology, Dankook University, Yongin 448-701, Korea.
authors.forename: Jonghoe
authors.initials: J
authors.lastname: Byun
mesh_terms: D000135:Acid Phosphatase / Q000235:genetics*; D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000494:pharmacology*; D001665:Binding Sites; D045744:Cell Line, Tumor; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D008297:Male; D008958:Models, Molecular; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000235:genetics; D052156:SELEX Aptamer Technique
chemical_list: D052157:Aptamers, Nucleotide; D000135:Acid Phosphatase; C428439:prostatic acid phosphatase
keywords: RNA aptamer; biomarker; prostate cancer; prostatic acid phosphatase