Identification and expression of troponin T, a new marker on the surface of cultured tumor endothelial cells by aptamer ligand

Cancer Medicine

Mst. Naznin Ara³  Mamoru Hyodo³  Noritaka Ohga¹  Kosuke Akiyama¹  Kyoko Hida¹  Yasuhiro Hida⁴  Nobuo Shinohara² 

[1] Division of Vascular Biology, Graduate School of Dental Medicine, Hokkaido University, Sapporo, Hokkaido, Japan;Department of Urology, Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido, Japan;Laboratory of Innovative Nanomedicine, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Hokkaido, Japan;Department of Cardiovascular and Thoracic Surgery, Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido, Japan

关键词: Biomarker;    DNA aptamer;    MALDI‐TOF‐mass spectroscopy;    mTECs;    troponin T;   

DOI  :  10.1002/cam4.260

来源: Wiley

【 摘 要 】

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 (K_d = 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.

【 授权许可】

CC BY   
© 2014 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.