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Telomerase as Biomarker for Oral Cancer

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Biomarkers in Cancer

Abstract

Telomerase is a ribonucleoprotein enzyme that extends telomere DNA located in the chromosomal termini. Telomerase is known to specifically express in cancer cells, and telomerase activity was found in more than 80 % of cancer patients. Therefore, telomerase may represent a promising cancer biomarker and therapeutic target. The telomerase repeat amplification protocol (TRAP) assay is a polymerase chain reaction (PCR)-based method to detect telomerase activity. In this assay, the telomerase substrate (TS) primer is elongated and its extended DNA is amplified by PCR. Telomerase activity is evaluated by a ladder of bands differing by 6 bp after gel electrophoresis of the PCR products. Recently, non-PCR-based methods to detect telomerase activity have been reported by many researchers. Electrochemical telomerase assay (ECTA), which was developed by Takenaka’s group, is a simple and rapid PCR-free method to detect telomerase activity. ECTA consists of a TS primer-immobilized electrode and ferrocenylnaphthalene diimide derivative as a tetraplex binder. Takenaka’s group compared the efficacy of ECTA and TRAP methods in detecting telomerase activity in oral cancer screening. Telomerase activity was observed in 90 % and 30 % of oral cancer tissues and exfoliative cells using the TRAP method, respectively, whereas the ECTA method detected telomerase activity in 90 % and 85 % of oral cancer tissues and exfoliative cells, respectively. These findings suggested that the ECTA method is useful for oral cancer screening because exfoliative cells can be easily obtained by scraping the inside of the mouth. Because telomerase activity is specific to cancer cells, ligands that inhibit telomerase activity may show promise as anticancer drugs. ECTA has also been used to estimate the inhibitory activity of telomerase ligands in cancer cells by determining TS primer elongation in a concentration-dependent manner. The ECTA method was used to screen for the telomerase inhibitory activity of 10 ligands and demonstrated not only the inhibitory activity of the ligands but also their mechanisms of action. The drugs having non-elongating capability directly inhibited telomerase. On the other hand, another set of drugs that can elongate at a 24-mer expansion indirectly inhibited telomerase access by binding and stabilizing the tetraplex structures.

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Abbreviations

Ca9-22:

Human Gingival Squamous Cell Carcinoma Cell Lines

ECTA:

Electrochemical Telomerase Assay

EtBr:

Ethidium Bromide

FND:

Ferrocenylnaphthalene Diimide

HSC-2:

Human Oral Squamous Cell Carcinoma Cell Lines

HSC-3:

Human Tongue Squamous Cell Carcinoma Cell Lines

hTERT:

Human Telomerase Reverse Transcriptase

IC50 :

Half maximal (50 %) Inhibitory Concentration

Inhibitor III:

Hexameric Phosphorothioate Oligonucleotide, 5′-d(TTAGGG)-3′

Inhibitor V:

2,6-bis[3-(N-piperidino)propionamido]anthracene-9,10-dione

PCR:

Polymerase Chain Reaction

PIPER:

N,N′-bis [2-(1-piperidino)ethyl]-3,4,9,10-tetracarboxylic diimide

Q:

Charge Quantity

SAS:

Human Tongue Squamous Cell Carcinoma Cell Line

SPR:

Surface Plasmon Resonance

SWV:

Square Wave Voltammetry

TBM:

3,3′,5,5′-Tetramethylbenzidine

TER:

Telomerase RNA Component

TERT:

Telomerase Reverse Transcriptase

TMPyP4:

5,10,15,20-tetra-(N-methyl-4-pyridyl)porphine

TND:

Tri Naphthalene Diimide

TRAP:

Telomerase Repeat Amplification protocol

TS:

Primer Telomerase Substrate Primer

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Authors and Affiliations

  1. Department of Applied Chemistry, Research Center for Bio-microsensing Technology, Kyushu Institute of Technology, Kitakyushu, Fukuoka, Japan

    Shigeori Takenaka & Shinobu Sato

Authors
  1. Shigeori Takenaka
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  2. Shinobu Sato
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Correspondence to Shigeori Takenaka .

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Editors and Affiliations

  1. Faculty of Life Sciences & Medicine King’s College London, Department of Nutrition and Dietetics Division of Diabetes & Nutritional Scien, London, United Kingdom

    Victor R. Preedy

  2. Faculty of Science & Technology, Department of Biomedical Sciences, University of Westminster, London, United Kingdom

    Vinood B. Patel

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Takenaka, S., Sato, S. (2015). Telomerase as Biomarker for Oral Cancer. In: Preedy, V., Patel, V. (eds) Biomarkers in Cancer. Biomarkers in Disease: Methods, Discoveries and Applications. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-7681-4_8

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