使用基于定量PCR的端粒重复序列扩增协议（qTRAP）评估哺乳动物细胞中的端粒酶活性。

Assessing Telomerase Activities in Mammalian Cells Using the Quantitative PCR-Based Telomeric Repeat Amplification Protocol (qTRAP).

作者信息

Jiang Shuai, Tang Mengfan, Xin Huawei, Huang Junjiu

机构信息

Key Laboratory of Gene Engineering of the Ministry of Education, SYSU-BCM Joint Center for Biomedical Sciences and Institute of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University, No. 135, Xingang West Road, Guangzhou, China.

Department of Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd, Houston, TX, USA.

出版信息

Methods Mol Biol. 2017;1587:95-101. doi: 10.1007/978-1-4939-6892-3_9.

DOI:10.1007/978-1-4939-6892-3_9

PMID:28324501

Abstract

Telomerase expression and activity appear elevated in >80% of human cancers. The activity of the telomerase may serve as a diagnostic marker for malignancy, and an indicator of the proliferative potential of somatic and stem cells. The telomeric repeat amplification protocol (TRAP) is a sensitive and accurate PCR-based assay for telomerase detection and measurement. Here, we describe a quantitative PCR-based TRAP assay (qTRAP) that is more convenient and amenable to high-throughput applications compared to traditional gel-based TRAP assays. qTRAP can not only facilitate drug screening processes for compounds that regulate telomerase activities but also enable the measurement of total telomerase activities of cultured cells or clinical specimens; the latter should prove particularly valuable to investigators of malignancies and diseases that are associated with telomerase and telomere dysfunction.

摘要

在超过80%的人类癌症中，端粒酶的表达和活性似乎有所升高。端粒酶的活性可作为恶性肿瘤的诊断标志物，以及体细胞和干细胞增殖潜力的指标。端粒重复序列扩增法（TRAP）是一种基于聚合酶链反应（PCR）的灵敏且准确的端粒酶检测和测量方法。在此，我们描述了一种基于定量PCR的TRAP检测法（qTRAP），与传统的基于凝胶的TRAP检测法相比，它更便捷且适用于高通量应用。qTRAP不仅有助于对调节端粒酶活性的化合物进行药物筛选，还能测量培养细胞或临床标本中的总端粒酶活性；对于研究与端粒酶和端粒功能障碍相关的恶性肿瘤和疾病的研究人员而言，后者应具有特别重要的价值。