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Targeted therapy of human laryngeal squamous cell carcinoma in vitro by antisense oligonucleotides directed against telomerase reverse transcriptase mRNA

Published online by Cambridge University Press:  16 March 2006

Zezhang Tao ,
Shiming Chen ,
Zhanyuan Wu ,
Bokui Xiao ,
Jianfeng Liu  and
Wei Hou
Zezhang Tao
Affiliation:
Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, People’s Republic of China
Shiming Chen
Affiliation:
Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, People’s Republic of China
Zhanyuan Wu
Affiliation:
Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, People’s Republic of China
Bokui Xiao
Affiliation:
Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, People’s Republic of China
Jianfeng Liu
Affiliation:
Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, People’s Republic of China
Wei Hou
Affiliation:
Virus Institute, College of Medicine, Wuhan University, Wuhan, People’s Republic of China
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Abstract

A number of different approaches have been developed to inhibit telomerase activity in human cancer cells. In this study, the effect of antisense oligonucleotides (ODNs) by targeting human telomerase reverse transcriptase (hTERT) mRNA in a laryngeal cancer cell line (Hep-2) was investigated. A 20mer antisense oligodeoxynucleotide targeting the most open part of hTERT mRNA (anti-hTERT) and a mismatched control sequence were synthesized. Cells were treated daily with oligonucleotides for up to 72 hours. hTERT mRNA expression was measured by the reverse transcription polymerase chain reaction (RT-PCR) assay; telomerase activity by the telomerase PCR ELISA assay kit (TRAP; Boehringer Mannheim, GmbH, Mannheim, Germany). Cell viability after administration of ODNs was determined using the MTT assay. Morphological changes were examined by haematoxylin and eosin staining. The cell cycle was analyzed using flow cytometry.

It was found that antisense treatment induced a decrease in hTERT mRNA expression, telomerase activity, cell growth rate, cell viability, and an increase in apoptosis. The results suggest that inhibition of telomerase activity in Hep-2 cells by short-term antisense treatment against the mRNA of hTERT results in apoptotic cell death. The treatment with anti-hTERT may be useful as a treatment modality for laryngeal squamous carcinoma.

Keywords

TelomeraseAntisense OligonucleotideLarynxSquamous Cell Carcinoma
Type
Research Article
Information
The Journal of Laryngology & Otology , Volume 119 , Issue 2 , February 2005 , pp. 92 - 96
Copyright
© 2005 Royal Society of Medicine Press

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