title: Identification and expression of troponin T, a new marker on the surface of cultured tumor endothelial cells by aptamer ligand.
abstract: The identification of a specific biomarker involves the development of new clinical diagnostic tools, and an in-depth understanding of the disease at the molecular level. When new blood vessels form in tumor cells, endothelial cell production is induced, a process that plays a key role in disease progression and metastasis to distinct organs for solid tumor types. The present study reports on the identification of a new biomarker on primary cultured mouse tumor endothelial cells (mTECs) using our recently developed high-affinity DNA aptamer AraHH001 (Kd = 43 nmol/L) assisted proteomics approach. We applied a strategy involving aptamer-facilitated biomarker discovery. Biotin-tagged AraHH001 was incubated with lysates of mTECs and the aptamer-proteins were then conjugated with streptavidin magnetic beads. Finally, the bound proteins were separated by sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. We identified troponin T via matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry, the molecular target of aptamer AraHH001, and its presence was confirmed by measuring mRNA, protein levels, western blot, immunostaining, a gel shift assay of AraHH001 with troponin T. We first report here on the discovery of troponin T on mTECs, a promising and interesting diagnostic tool in the development of antiangiogenic therapy techniques the involves the targeting of the tumor vasculature.
authors.affiliation: Laboratory of Innovative Nanomedicine, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita 12, Nishi 6, Kita-ku, Sapporo, Hokkaido, 060-0812, Japan.
authors.forename: Mst Naznin
authors.initials: MN
authors.lastname: Ara
authors.forename: Mamoru
authors.initials: M
authors.lastname: Hyodo
authors.forename: Noritaka
authors.initials: N
authors.lastname: Ohga
authors.forename: Kosuke
authors.initials: K
authors.lastname: Akiyama
authors.forename: Kyoko
authors.initials: K
authors.lastname: Hida
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Hida
authors.forename: Nobuo
authors.initials: N
authors.lastname: Shinohara
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry; D052158:Aptamers, Peptide; D014408:Biomarkers, Tumor / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D051379:Mice; D052156:SELEX Aptamer Technique; D013194:Staining and Labeling; D020107:Troponin T / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D052157:Aptamers, Nucleotide; D052158:Aptamers, Peptide; D014408:Biomarkers, Tumor; D020107:Troponin T
keywords: Biomarker; DNA aptamer; MALDI-TOF-mass spectroscopy; mTECs; troponin T