Assessment Effects of Resveratrol on Human Telomerase Reverse Transcriptase Messenger Ribonucleic Acid Transcript in Human Glioblastoma

Mirzazadeh, Azin; Kheirollahi, Majid; Farashahi, Ehsan; Sadeghian-Nodoushan, Fatemeh; Sheikhha, Mohammad Hasan; Aflatoonian, Behrouz

Assessment Effects of Resveratrol on Human Telomerase Reverse Transcriptase Messenger Ribonucleic Acid Transcript in Human Glioblastoma

Document Type : Original Article

Authors

¹ Department of Medical Genetics, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

² Department of Medical Genetics, Pediatrics Inherited Diseases Research Center, Research Institute for Primordial Prevention of Noncommunicable Disease School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

³ Department of Medical Genetics; Research and Clinical Centre for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

⁴ Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

Abstract

Background: Glioblastoma (GBM) is the most common and aggressive brain tumor, which has a poor prognosis despite the advent of different therapeutic strategies. There are numerous molecular biomarkers to contribute diagnosis, prognosis, and prediction of response to the current therapy in GBM. One of the most important markers that are potentially valuable is immortalization-specific or immortalization-associated marker named “hTERT messenger ribonucleic acid (mRNA)” the key subunit of telomerase enzyme, which is expressed in more than 85% of cancer cells, in spite of the majority of normal somatic cells. In this study, we investigated the effects of resveratrol (RSV) on this mRNA marker level, leading to cancer progression. Materials and Methods: U-87MG cell line was obtained from Pasteur Institute of Iran and treated with various concentrations of 0–160 μg/mL of RSV and at different time points (24, 48, and 72 h). To evaluate viability of U-87MG cells, standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed. Real-time polymerase chain reaction (RT-PCR) was used for comparative and quantitative assessment of human telomerase reverse transcriptase (hTERT) mRNA copy number versus control–untreated group. Results: The results of our investigation suggested that RSV effectively inhibited cell growth and caused cell death in dose-dependent (P < 0.05) and not in time-dependent manner (P > 0.05), in vitro. Interestingly, quantitative RT-PCR analysis demonstrated that at half inhibition concentration, RSV dramatically decreased mRNA expression of hTERT, the catalytic subunit of telomerase enzyme, which leads to prevention of cell division and tumor progression. Conclusion: With regard to downregulation of this immortalization-associated marker, RSV may potentially be used as a therapeutic agent against GBM.

Keywords

References

1.	Bleeker FE, Molenaar RJ, Leenstra S. Recent advances in the molecular understanding of glioblastoma. J Neurooncol 2012;108:11-27.
2.	Zamin LL, Filippi-Chiela EC, Dillenburg-Pilla P, Horn F, Salbego C, Lenz G. Resveratrol and quercetin cooperate to induce senescence-like growth arrest in C6 rat glioma cells. Cancer Sci 2009;100:1655-62.
3.	Van Meir EG, Hadjipanayis CG, Norden AD, Shu HK, Wen PY, Olson JJ. Exciting new advances in neuro-oncology: The avenue to a cure for malignant glioma. CA Cancer J Clin 2010;60:166-93.
4.	Galli R, Binda E, Orfanelli U, Cipelletti B, Gritti A, De Vitis S, et al. Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma. Cancer Res 2004;64:7011-21.
5.	McNamara MG, Sahebjam S, Mason WP. Emerging biomarkers in glioblastoma. Cancers (Basel) 2013;5:1103-19.
6.	Snijders PJ, Breuer RH, Sutedja TG, Egging M, Voorhorst FJ, Steenbergen RD, et al. Elevated hTERT mRNA levels: A potential determinant of bronchial squamous cell carcinoma (in situ). Int J Cancer 2004;109:412-7.
7.	Athar M, Back JH, Tang X, Kim KH, Kopelovich L, Bickers DR, et al. Resveratrol: A review of preclinical studies for human cancer prevention. Toxicol Appl Pharmacol 2007;224:274-83.
8.	Kala R, Tollefsbol TO, Li Y. Potential of Resveratrol in Inhibiting Cancer and Slowing Aging. Journal of Nutrition and Food Sciences. 2012 Nov 27;2015.
9.	Jiang H, Zhang L, Kuo J, Kuo K, Gautam SC, Groc L, et al. Resveratrol-induced apoptotic death in human U251 glioma cells. Mol Cancer Ther 2005;4:554-61.
10.	Steinbach JP, Weller M. Apoptosis in gliomas: Molecular mechanisms and therapeutic implications. J Neurooncol 2004;70:245-54.
11.	Yang YP, Chang YL, Huang PI, Chiou GY, Tseng LM, Chiou SH, et al. Resveratrol suppresses tumorigenicity and enhances radiosensitivity in primary glioblastoma tumor initiating cells by inhibiting the STAT3 axis. J Cell Physiol 2012;227:976-93.
12.	George J, Banik NL, Ray SK. Knockdown of hTERT and concurrent treatment with interferon-gamma inhibited proliferation and invasion of human glioblastoma cell lines. Int J Biochem Cell Biol 2010;42:1164-73.
13.	Sprouse AA, Steding CE, Herbert BS. Pharmaceutical regulation of telomerase and its clinical potential. J Cell Mol Med 2012;16:1-7.
14.	Boldrini L, Pistolesi S, Gisfredi S, Ursino S, Ali G, Pieracci N, et al. Telomerase activity and hTERT mRNA expression in glial tumors. Int J Oncol 2006;28:1555-60.
15.	Pezzuto JM. Resveratrol as an inhibitor of carcinogenesis. Pharm Biol 2008;46:443-573.
16.	Lanzilli G, Fuggetta MP, Tricarico M, Cottarelli A, Serafino A, Falchetti R, et al. Resveratrol down-regulates the growth and telomerase activity of breast cancer cells in vitro. Int J Oncol 2006;28:641-8.
17.	Fuggetta MP, Lanzilli G, Tricarico M, Cottarelli A, Falchetti R, Ravagnan G, et al. Effect of resveratrol on proliferation and telomerase activity of human colon cancer cells in vitro. J Exp Clin Cancer Res 2006;25:189-93.
18.	Olaussen KA, Dubrana K, Domont J, Spano JP, Sabatier L, Soria JC. Telomeres and telomerase as targets for anticancer drug development. Crit Rev Oncol Hematol 2006;57:191-214.
19.	Shervington A, Patel R. Differential hTERT mRNA processing between young and older glioma patients. FEBS Lett 2008;582:1707-10.
20.	Agrawal A, Dang S, Gabrani R. Recent patents on anti-telomerase cancer therapy. Recent Pat Anticancer Drug Discov 2012;7:102-17.
21.	Fujiwara M, Okayasu I, Takemura T, Tanaka I, Masuda R, Furuhata Y, et al. Telomerase activity significantly correlates with chromosome alterations, cell differentiation, and proliferation in lung adenocarcinomas. Mod Pathol 2000;13:723-9.
22.	Nakatani K, Yoshimi N, Mori H, Yoshimura S, Sakai H, Shinoda J, et al. The significant role of telomerase activity in human brain tumors. Cancer 1997;80:471-6.
23.	Kheirollahi M, Mehrazin M, Kamalian N, Mohammadi-asl J, Mehdipour P. Telomerase Activity in Human Brain Tumors: Astrocytoma and Meningioma. Cell Mol Neurobiol 2013 33:569–574.

Advanced Biomedical Research