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: Mst Naznin Ara, Mamoru Hyodo, Noritaka Ohga, Kosuke Akiyama, Kyoko Hida, Yasuhiro Hida, Nobuo Shinohara, Hideyoshi Harashima
• Journal: Cancer medicine (Cancer Med) Vol. 3 Issue 4 Pg. 825-34 (Aug 2014) ISSN: 2045-7634 [Electronic] United States
• PMID: 24810801 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

Chemical References

• Aptamers, Nucleotide
• Aptamers, Peptide
• Biomarkers, Tumor
• Troponin T

Topics

• Animals
• Aptamers, Nucleotide (chemistry)
• Aptamers, Peptide
• Biomarkers, Tumor (metabolism)
• Endothelial Cells (metabolism)
• Humans
• Mice
• SELEX Aptamer Technique
• Staining and Labeling
• Troponin T (metabolism)
• Tumor Cells, Cultured