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Inhibitory effect of HuR gene small interfering RNA segment on laryngeal carcinoma Hep-2 cell growth

Published online by Cambridge University Press:  02 June 2010

Z Shen ,
D Ye ,
X Zhang ,
Z Jiang ,
B Xiao  and
J Guo
Z Shen*
Affiliation:
Department of Otorhinolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, China Institute of Biochemistry and Molecular Biology, Ningbo University School of Medicine, China
D Ye
Affiliation:
Department of Otorhinolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, China Institute of Biochemistry and Molecular Biology, Ningbo University School of Medicine, China
X Zhang
Affiliation:
Department of Otorhinolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, China Institute of Biochemistry and Molecular Biology, Ningbo University School of Medicine, China
Z Jiang
Affiliation:
Institute of Biochemistry and Molecular Biology, Ningbo University School of Medicine, China
B Xiao
Affiliation:
Institute of Biochemistry and Molecular Biology, Ningbo University School of Medicine, China
J Guo
Affiliation:
Institute of Biochemistry and Molecular Biology, Ningbo University School of Medicine, China
*
Address for correspondence: Zhisen Shen, M.D., Department of Otorhinolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, Ningbo, 315000, China. Tel.: +86-13906687216 Fax: +86-574-87392232 E-mail: szs7216@sina.com
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Abstract

Objectives:

To investigate the effect of the HuR gene on laryngeal carcinoma Hep-2 cell growth, and to analyse correlations between the HuR, cyclooxygenase-2 and survivin genes.

Study design:

Experiment study.

Setting:

Department of Otolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, Ningbo, a tertiary care centre in China.

Methods:

Copies of a small interfering RNA segment directed against the HuR gene were transfected into Hep-2 cells using LipofectamineTM 2000. The effect of the small interfering RNA segment on Hep-2 cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Changes in the expression of the HuR, cyclooxygenase-2 and survivin genes were detected by semi-quantitative reverse transcription polymerase chain reaction analysis. Concentrations of the HuR, cyclooxygenase-2 and survivin proteins were evaluated using Western blotting.

Results:

Expression of the HuR, cyclooxygenase-2 and survivin genes, as indicated by messenger RNA and protein levels, was suppressed by the HuR gene small interfering RNA segment in a dose-dependent manner. The proliferation indices of all treated groups were significanlty lower than those of control groups (p < 0.05).

Conclusions:

Impairment of HuR gene expression, using interfering RNA technology, can significantly suppress Hep-2 cell proliferation and induce apoptosis. The HuR gene may be an effective target for gene therapy in patients with laryngeal carcinoma.

Keywords

Laryngeal NeoplasmsCarcinomaSurvivin ProteinHuman GeneHuR Gene
Type
Main Articles
Information
The Journal of Laryngology & Otology , Volume 124 , Issue 11 , November 2010 , pp. 1183 - 1189
Copyright
Copyright © JLO (1984) Limited 2010

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References

1 Chu, EA, Kim, YJ. Laryngeal cancer: diagnosis and preoperative work-up. Otolaryngol Clin North Am 2008;41:673–95CrossRefGoogle ScholarPubMed
2 Benasso, M, Bonelli, L, Numico, G, Corvò, R, Sanguineti, G, Rosso, R et al. Treatment with cisplatin and fluorouracil alternating with radiation favourably affects prognosis of inoperable squamous cell carcinoma of the head and neck: results of a multivariate analysis on 273 patients. Ann Oncol 1997;8:773–9CrossRefGoogle ScholarPubMed
3 Do, SI, Araujo, ES, Kalil, RK, Bacchini, P, Bertoni, F, Unni, KK et al. Expression of embryonic lethal abnormal vision (ELAV)-like protein HuR and cyclooxygenase-2 (COX-2) in Ewing sarcoma. Tumori 2008;94:347–50CrossRefGoogle Scholar
4 Ma, WJ, Cheng, S, Campbell, C, Wright, A, Furneaux, H. Cloning and characterization of HuR, a ubiquitously expressed Elav-like protein. J Biol Chem 1996;271:8144–51CrossRefGoogle Scholar
5 Ross, J. mRNA stability in mammalian cells. Microbiol Rev 1995;59:423–50CrossRefGoogle ScholarPubMed
6 Denkert, C, Weichert, W, Winzer, KJ, Müller, BM, Noske, A, Niesporek, S et al. Expression of the ELAV-like protein HuR is associated with higher tumor grade and increased cyclooxygenase-2 expression in human breast carcinoma. Clin Cancer Res 2004;10:5580–6CrossRefGoogle ScholarPubMed
7 Denkert, C, Koch, I, von Keyserlingk, N, Noske, A, Niesporek, S, Dietel, M et al. Expression of the ELAV-like protein HuR in human colon cancer: association with tumor stage and cyclooxygenase-2. Mod Pathol 2006;19:1261–9CrossRefGoogle ScholarPubMed
8 Erkinheimo, TL, Sivula, A, Lassus, H, Heinonen, M, Furneaux, H, Haglund, C et al. Cytoplasmic HuR expression correlates with epithelial cancer cell but not with stromal cell cyclooxygenase-2 expression in mucinous ovarian carcinoma. Gynecol Oncol 2005;99:14–19CrossRefGoogle Scholar
9 Mrena, J, Wiksten, JP, Thiel, A, Kokkola, A, Pohjola, L, Lundin, J et al. Cyclooxygenase-2 is an independent prognostic factor in gastric cancer and its expression is regulated by the messenger RNA stability factor HuR. Clin Cancer Res 2005;11:7362–8CrossRefGoogle ScholarPubMed
10 Nabors, LB, Gillespie, GY, Harkins, L, King, PH. HuR, a RNA stability factor, is expressed in malignant brain tumors and binds to adenine- and uridine-rich elements within the 3′ untranslated regions of cytokine and angiogenic factor mRNAs. Cancer Res 2001;61:2154–61Google Scholar
11 Cho, NP, Han, HS, Soh, Y, Lee, KY, Son, HJ. Cytoplasmic HuR over-expression is associated with increased cyclooxygenase-2 expression in laryngeal squamous cell carcinomas. Pathology 2007;39:545–50CrossRefGoogle ScholarPubMed
12 Dubois, RN, Abramson, SB, Crofford, L, Gupta, RA, Simon, LS, Van De Putte, LB et al. Cyclooxygenase in biology and disease. FASEB J 1998;12:1063–73CrossRefGoogle ScholarPubMed
13 Gupta, RA, Dubois, RN. Colorectal cancer prevention and treatment by inhibition of cyclooxygenase-2. Nat Rev Cancer 2001;1:11–21CrossRefGoogle ScholarPubMed
14 Kolev, Y, Uetake, H, Iida, S, Ishikawa, T, Kawano, T, Sugihara, K. Prognostic significance of VEGF expression in correlation with COX-2, microvessel density, and clinicopathological characteristics in human gastric carcinoma. Ann Surg Oncol 2007;14:2738–47CrossRefGoogle ScholarPubMed
15 Cho, SJ, Kim, N, Kim, JS, Jung, HC, Song, IS. The anti-cancer effect of COX-2 inhibitors on gastric cancer cells. Dig Dis Sci 2007;52:1713–21CrossRefGoogle ScholarPubMed
16 Lu, B, Mu, Y, Cao, C, Zeng, F, Schneider, S, Tan, J et al. Survivin as a therapeutic target for radiation sensitization in lung cancer. Cancer Res 2004;64:2840–5CrossRefGoogle ScholarPubMed
17 Kawasaki, H, Toyoda, M, Shinohara, H, Okuda, J, Watanabe, I, Yamamoto, T et al. Expression of survivin correlates with apoptosis, proliferation, and angiogenesis during human colorectal tumorigenesis. Cancer 2001;91:2026–323.0.CO;2-E>CrossRefGoogle ScholarPubMed
18 Kato, J, Kuwabara, Y, Mitani, M, Shinoda, N, Sato, A, Toyama, T et al. Expression of survivin in esophageal cancer: correlation with the prognosis and response to chemotherapy. Int J Cancer 2001;95:92–53.0.CO;2-9>CrossRefGoogle ScholarPubMed
19 Yin, RT, Peng, ZL, Yang, KX, Kang, DY. Survivin expression in ovarian carcinoma [in Chinese]. Sichuan Da Xue Xue Bao Yi Xue Ban 2006;37:215–17, 233Google ScholarPubMed
20 Wang, Y, Kong, W, Xiong, X, Zhang, S, Sun, D. The expressions of survivin and VEGF in squamous cell carcinoma of larynx and the correlation between the two marks [in Chinese]. Lin Chuang Er Bi Yan Hou Ke Za Zhi 2005;19:838–41Google ScholarPubMed
21 Fan, LF, Dong, WG, Jiang, CQ, Qian, Q, Yu, QF. Role of hypoxia-inducible factor-1 alpha and survivin in colorectal carcinoma progression. Int J Colorectal Dis 2008;23:1057–64CrossRefGoogle ScholarPubMed
22 Nassar, A, Sexton, D, Cotsonis, G, Cohen, C. Survivin expression in breast carcinoma: correlation with apoptosis and prognosis. Appl Immunohistochem Mol Morphol 2008;16:221–6CrossRefGoogle ScholarPubMed
23 Liu, L, Zhang, M, Zou, P. Expression of PLK1 and survivin in diffuse large B-cell lymphoma. Leuk Lymphoma 2007;48:2179–83CrossRefGoogle ScholarPubMed
24 Knutsen, A, Adell, G, Sun, XF. Survivin expression is an independent prognostic factor in rectal cancer patients with and without preoperative radiotherapy. Int J Radiat Oncol Biol Phys 2004;60:149–55CrossRefGoogle ScholarPubMed
25 Sachs, AB. Messenger RNA degradation in eukaryotes. Cell 1993;74:413–21CrossRefGoogle ScholarPubMed
26 Garneau, NL, Wilusz, J, Wilusz, CJ. The highways and byways of mRNA decay. Nat Rev Mol Cell Biol 2007;8:113–26CrossRefGoogle ScholarPubMed
27 Sengupta, S, Jang, BC, Wu, MT, Paik, JH, Furneaux, H, Hla, T. The RNA-binding protein HuR regulates the expression of cyclooxygenase-2. J Biol Chem 2003;278:25227–33CrossRefGoogle ScholarPubMed
28 Athanassiadou, P, Grapsa, D, Athanassiades, P, Gonidi, M, Athanassiadou, AM, Tsipis, A et al. The prognostic significance of COX-2 and survivin expression in ovarian cancer. Pathol Res Pract 2008;204:241–9CrossRefGoogle ScholarPubMed
29 Li, S, Tong, Q, Zhang, W, Wang, Q, Chen, Z, Wu, Q. Mechanism of growth inhibitory effects of cyclooxygenase-2 inhibitor-NS398 on cancer cells. Cancer Invest 2008;26:333–7CrossRefGoogle ScholarPubMed
30 Erkanli, S, Bolat, F, Kayaselcuk, F, Demirhan, B, Kuscu, E. COX-2 and survivin are overexpressed and positively correlated in endometrial carcinoma. Gynecol Oncol 2007;104:320–5CrossRefGoogle ScholarPubMed
31 Kozaki, K, Koshikawa, K, Tatematsu, Y, Miyaishi, O, Saito, H, Hida, T et al. Multi-faceted analyses of a highly metastatic human lung cancer cell line NCI-H460-LNM35 suggest mimicry of inflammatory cells in metastasis. Oncogene 2001;20:4228–34CrossRefGoogle ScholarPubMed
32 Krysan, K, Merchant, FH, Zhu, L, Dohadwala, M, Luo, J, Lin, Y et al. COX-2-dependent stabilization of survivin in non-small cell lung cancer. FASEB J 2004;18:206–8CrossRefGoogle ScholarPubMed
33 Muchmore, SW, Chen, J, Jakob, C, Zakula, D, Matayoshi, ED, Wu, W et al. Crystal structure and mutagenic analysis of the inhibitor-of-apoptosis protein survivin. Mol Cell 2000;6:173–82CrossRefGoogle ScholarPubMed
34 Sheng, H, Shao, J, Morrow, JD, Beauchamp, RD, DuBois, RN. Modulation of apoptosis and Bcl-2 expression by prostaglandin E2 in human colon cancer cells. Cancer Res 1998;58:362–6Google ScholarPubMed