Analytical Biochemistry 2011-10-01

5-Ethynyl-2'-deoxycytidine as a new agent for DNA labeling: detection of proliferating cells.

Dezhong Qu, Guoxing Wang, Zhe Wang, Li Zhou, Weilin Chi, Shujie Cong, Xiaoshuai Ren, Peizhou Liang, Biliang Zhang

文献索引：Anal. Biochem. 417 , 112-121, (2011)

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摘要

The labeling of newly synthesized DNA in cells to identify cell proliferation is an important experimental technique. The most accurate methods incorporate [(3)H]thymidine or 5-bromo-2'-deoxyruidine (BrdU) into dividing cells during S phase, which is subsequently detected by autoradiography or immunohistochemistry, directly measuring the newly synthesized DNA. Recently, a novel method was developed to detect DNA synthesis in proliferating cells based on a novel thymidine analog, 5-ethynyl-2'-deoxyuridine (EdU). EdU is incorporated into DNA and subsequently detected with a fluorescent azide via "click" chemistry. This novel technique is highly sensitive and does not require DNA denaturation. However, it was also found that EdU exhibits time-dependent inhibition effects on cell growth. Therefore, here we report a novel deoxycytidine analog, 5-ethynyl-2'-deoxycytidine (EdC), that can be used to detect DNA synthesis in vitro and in vivo at a similar sensitivity level compared with EdU. Furthermore, the EdC-induced cytotoxicity is much less than that of EdU when combined with thymidine. This will be a potential application for the long-term detection of proliferating cells.Copyright © 2011 Elsevier Inc. All rights reserved.