Zhao Yong, Shay Jerry W, Wright Woodring E
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Methods Mol Biol. 2011;735:63-75. doi: 10.1007/978-1-61779-092-8_7.
Telomerase is present in most human cancers, and proliferative stem cells including germline cells. Telomerase plays an essential role in tumorigenesis by maintaining/elongating telomeric DNA, and thus preventing the telomere shortening that results in replicative senescence. Understanding telomerase action in vivo has important implication for both cancer and aging, but there are not robust methods for monitoring telomerase action. By combining a series of cell biological and biochemical approaches, and taking advantage of the enzyme DSN that specifically cuts double-stranded DNA and releases the telomeric overhangs, we have developed a method to monitor telomerase action during one cell cycle. Here, we describe this method using HeLa carcinoma cells as an example.
端粒酶存在于大多数人类癌症以及包括生殖细胞在内的增殖性干细胞中。端粒酶通过维持/延长端粒DNA在肿瘤发生过程中发挥重要作用，从而防止导致复制性衰老的端粒缩短。了解端粒酶在体内的作用对癌症和衰老研究都具有重要意义，但目前尚无可靠的方法来监测端粒酶的作用。通过结合一系列细胞生物学和生化方法，并利用能特异性切割双链DNA并释放端粒突出端的DSN酶，我们开发了一种在一个细胞周期内监测端粒酶作用的方法。在此，我们以HeLa癌细胞为例描述该方法。
