Hostname: page-component-586b7cd67f-t7fkt Total loading time: 0 Render date: 2024-11-24T06:33:15.576Z Has data issue: false hasContentIssue false

Short hairpin ribonucleic acid targeting the telomerase catalytic unit of messenger ribonucleic acid significantly limits the growth of laryngeal squamous cell carcinoma in nude mice

Published online by Cambridge University Press:  25 June 2007

Xu-Hong Zhou
Affiliation:
Department of Otolaryngology, Zhongnan Hospital, Wuhan University, Wuhan, PR China.
Shi-Ming Chen
Affiliation:
Department of Otolaryngology-Head & Neck Surgery, Renmin Hospital, Wuhan University, Wuhan, PR China
Dan Liu
Affiliation:
Department of Otolaryngology-Head & Neck Surgery, Renmin Hospital, Wuhan University, Wuhan, PR China
Yan Wang
Affiliation:
Department of Otolaryngology-Head & Neck Surgery, Renmin Hospital, Wuhan University, Wuhan, PR China
Bo-Kui Xiao
Affiliation:
Department of Otolaryngology-Head & Neck Surgery, Renmin Hospital, Wuhan University, Wuhan, PR China
Ze-Zhang Tao*
Affiliation:
Department of Otolaryngology-Head & Neck Surgery, Renmin Hospital, Wuhan University, Wuhan, PR China
*
Address for correspondence: Dr Ze-Zhang Tao, Department of Otolaryngology-Head & Neck Surgery, Renmin Hospital, Wuhan University, Wuhan 430060, PR China. Fax: +86 27 88043958, E-mail: [email protected]

Abstract

Objective:

Telomerase is an attractive molecular target because it is active in most malignant cells but undetectable in most normal somatic cells. Small, interfering ribonucleic acid segments have been shown to be effective tools for inhibiting the expression of a given gene within human cells. In the present study, we examined the effects of short hairpin ribonucleic acid expression vectors on the growth of laryngeal squamous cell carcinoma in nude mice, and we assessed potential side effects in these animals.

Methods:

Short hairpin ribonucleic acid expression vectors targeting the messenger ribonucleic acid of the telomerase catalytic unit were constructed and transfected into Hep-2 human laryngeal squamous cells carcinoma in nude mice. Apoptosis and telomerase catalytic unit expression within tumour cells were evaluated after treating with short hairpin ribonucleic acid. Peripheral blood was collected for haematological and biochemical analysis.

Results:

The findings demonstrated that short hairpin ribonucleic acid plasmids could inhibit tumour cell growth by 76.5 per cent, and that many tumour cells underwent necrotic or apoptotic cell death. There were no significant side effects of short hairpin ribonucleic acid on the heart, liver, kidney, spleen or blood system in this experimental model.

Conclusion:

These results indicated that the short hairpin ribonucleic acid expression vector targeted at the telomerase catalytic unit of messenger ribonucleic acid significantly inhibited the growth of laryngeal carcinoma in nude mice, with no significant side effects on the experimental animals.

Type
Main Article
Copyright
Copyright © JLO (1984) Limited 2007

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1 Shay, JW, Baccheter, S. A survey of telomerase activity in human cancer. Eur J Cancer 1997;33:787–91CrossRefGoogle ScholarPubMed
2 Masutomi, K, Yu, EY, Khurts, S, Ben-Porath, I, Currier, JL, Metz, GB et al. Telomerase maintains telomere structure in normal human cells. Cell 2003;114:241–53CrossRefGoogle ScholarPubMed
3 Luzar, B, Poljak, M, Marin, IJ, Fischinger, J, Gale, N. Quantitative measurement of telomerase catalytic subunit (hTERT) RNA in laryngeal squamous cell carcinomas. Anticancer Res 2001;21:4011–55Google ScholarPubMed
4 Curran, AJ, Gullane, PJ, Irish, J, MacMillan, C, Freeman, J, Kamel-Reid, S. Telomerase activity is upregulated in laryngeal squamous cell carcinoma. Laryngoscope 2000;110:391–6CrossRefGoogle ScholarPubMed
5 Feng, J, Funk, WD, Wang, SS, Weinrich, SL, Avilion, AA, Chiu, CP et al. The RNA component of human telomerase. Science 1995;269:1236–41CrossRefGoogle ScholarPubMed
6 Harrington, L, McPhail, T, Mar, V, Zhou, W, Oulton, R, Bass, MB et al. A mammalian telomerase-associated protein. Science 1997;275:973–7CrossRefGoogle ScholarPubMed
7 Nakayama, J, Saito, M, Nakamura, H, Matsuura, A, Ishikawa, F. TLP1: a gene encoding a protein component of mammalian telomerase is a novel member of WD repeats family. Cell 1997;88:875–84CrossRefGoogle ScholarPubMed
8 Meyerson, M, Counter, CM, Eaton, EN, Ellisen, LW, Steiner, P, Caddle, SD et al. hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization. Cell 1997;90:785–95CrossRefGoogle ScholarPubMed
9 Nakamura, TM, Morin, GB, Chapman, KB, Weinrich, SL, Andrews, WH, Lingner, J et al. Telomerase catalytic subunit homologs from fission yeast and human. Science 1997;277:955–9CrossRefGoogle ScholarPubMed
10 Kang, MK, Kameta, A, Shin, KH, Baluda, MA, Park, NH. Senescence occurs with hTERT repression and limited telomere shortening in human oral keratinocytes cultured with feeder cells. J Cell Physiol 2004;199:364–70CrossRefGoogle ScholarPubMed
11 Kyo, S, Inous, M. Complex regulatory mechanism of telomerase activity in normal and cancer cells: how can we apply them for cancer therapy? Oncogene 2002;21:688–97CrossRefGoogle ScholarPubMed
12 Mittal, A, Pate, MS, Wylie, RC, Tollefsbol, TO, Katiyar, SK. EGCG down-regulates telomerase in human breast carcinoma MCF-7 cells, leading to suppression of cell viability and induction of apoptosis. Int J Oncol 2004;24:703–10Google ScholarPubMed
13 Tao, Z, Chen, S, Wu, Z, Xiao, B, Liu, J, Hou, W. Targeted therapy of human laryngeal squamous cell carcinoma in vitro by antisense oligonucleotides directed against telomerase reverse transcriptase mRNA. J Laryngol Otol 2005;119:92–6CrossRefGoogle ScholarPubMed
14 Hannon, GJ. RNA interference. Nature 2002;418:244–51CrossRefGoogle ScholarPubMed
15 Liu, XD, Ma, SM, Liu, Y, Liu, SZ, Sehon, A. Short hairpin RNA and retroviral vector-mediated silencing of p53 in mammalian cells. Biochem Biophys Res Commun 2004;324:1173–8CrossRefGoogle ScholarPubMed
16 Montgomery, MK. RNA interference: historical overview and significance. Methods Mol Biol 2004;265:321Google ScholarPubMed
17 Fire, A, Xu, S, Montgomery, MK, Kostas, SA, Driver, SE, Mello, CC et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998;391:806–11CrossRefGoogle ScholarPubMed
18 Elbashir, SM, Harborth, J, Lendeckel, W, Yalcin, A, Weber, K, Tuschl, T. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 2001;411:494–8CrossRefGoogle ScholarPubMed
19 Paddison, PJ, Caudy, AA, Bernstein, E, Hannon, GJ, Conklin, DS. Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes Dev 2002;16:948–58CrossRefGoogle ScholarPubMed
20 Brummelkamp, TR, Bernards, R, Agami, R. A system for stable expression of short interfering RNAs in mammalian cells. Science 2002;296:550–3CrossRefGoogle ScholarPubMed
21 Chen, SM, Tao, ZZ, Hua, QQ, Liu, D, Chi, HM, Cai, Q. Inhibition of human telomerase reverse transcriptase in Hep-2 cells using short hairpin RNA expression vectors. Arch Otolaryngol Head Neck Surg 2006;132:200–5CrossRefGoogle ScholarPubMed
22 Zhang, DL, Zhang, YT, Yin, JJ, Zhao, BL. Oral administration of crataegus flavonoids protects against ischemia/reperfusion brain damage in gerbils. J Neurochem 2004;90:211–19CrossRefGoogle ScholarPubMed
23 Zhou, XD, Yu, JP, Liu, J, Luo, HS, Chen, HX, Yu, HG. Overexpression of cellular FLICE-inhibitory protein (FLIP) in gastric adenocarcinoma. Clin Sci (Lond) 2004;106:397405CrossRefGoogle ScholarPubMed
24 Simonsen, JL, Rosada, C, Serakinci, N, Justesen, J, Stenderup, K, Rattan, SI et al. Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells. Nat Biotechnol 2002;20:592–6CrossRefGoogle ScholarPubMed
25 Folini, M, Brambilla, C, Villa, R, Gandellini, P, Vignati, S, Paduano, F et al. Antisense oligonucleotide-mediated inhibition of hTERT, but not hTERC, induces rapid cell growth decline and apoptosis in the absence of telomere shortening in human prostate cancer cells. Eur J Cancer 2005;41:624–34CrossRefGoogle Scholar
26 Liu, Y, Snow, BE, Hande, MP, Baerlocher, G, Kickhoefer, VA, Yeung, D et al. The telomerase reverse transcriptase is limiting and necessary for telomerase function in vivo. Curr Biol 2000;10:1459–62CrossRefGoogle ScholarPubMed
27 Koyanagi, Y, Kobayashi, D, Yajima, T, Asanuma, K, Kimura, T, Sato, T et al. Telomerase activity is down regulated via decreases in hTERT mRNA but not TEP1 mRNA or hTERC during the differentiation of leukemic cells. Anticancer Res 2000;20:773–8Google ScholarPubMed