The suppression of manganese superoxide dismutase decreased the survival of human glioblastoma multiforme T98G cells
Background: Glioblastoma multiforme (GBM) is a primary malignant brain tumor which has poor prognosis. High incidence of oxidative stress-based therapy resistance could be related to the high antioxidant status of GBM cells. Our previous study has reported that manganese superoxide dismutase (MnSOD) antioxidant expression was significantly higher in high grade glioma than in low grade. The aim of this study was to analyze the impact of MnSOD suppression toward GBM cell survival.
Methods: This study is an experimental study using human glioblastoma multiforme T98G cell line. Suppression of MnSOD expression was performed using in vitro transfection MnSOD-siRNA. The MnSOD expression was analyzed by measuring the mRNA using real time RT-PCR, protein using ELISA technique, and specific activity of enzyme using inhibition of xantine oxidase. Concentration of reactive oxygen species (ROS) intracellular was determined by measuring superoxide radical and hydrogen peroxide. Cell survival was analyzed by measuring viability, proliferation, and cell apoptosis.
Results: In vitro transfection of MnSOD-siRNA suppressed the mRNA, protein, and specific activity of MnSOD. This treatment significantly increased the concentration of superoxide radical; however, it did not influence the concentration of hydrogen peroxide. Moreover, viability MnSOD-suppressing cell significantly decreased, accompanied by increase of cell apoptosis without affecting cell proliferation.
Conclusion: The suppression of MnSOD expression leads to decrease glioblastoma multiforme cell survival, which was associated to the increase of cell apoptotic.
Walid MS. Prognostic factor for long-term survival after glioblastoma. Perm J. 2008;12(4):45–8. https://doi.org/10.7812/TPP/08-027
Azzam EI, Jay-Gerin JP, Pain D. Ionizing radiation-induced metabolic oxidative stress and prolonged cell injury. Cancer Lett. 2012;327(1-2):48–60. https://doi.org/10.1016/j.canlet.2011.12.012
Halliwell B, Gutteridge JMC. Free radical in biology and medicine. 4th ed. London: Oxford University Press; 2007. p. 30–74.
Dhar SK, Tangpong J, Chaiswing L, Oberley TD, St. Clair DK. Manganese superoxide dismutase is a p53-regulated gene that switches cancers between early and advanced staged. Cancer Res. 2011;71(21):6684–95. https://doi.org/10.1158/0008-5472.CAN-11-1233
Kattan J, Minig V, Leroy P, Dauça M, Becuwe P. Role of manganese superoxide dismutase on growth and invasive properties of human estrogen-independent breast cancer cells. Breast Cancer Res Treat. 2008;108(2):203–15. https://doi.org/10.1007/s10549-007-9597-5
Järvelä S, Bragge H, Paunu N, Järvelä T, Paljärvi H, Kalimo H, et al. Antioxidant enzymes in oligodendroglial brain tumors: association with proliferation, apoptotic activity and survival. J Neurooncol. 2006;77(2):131–40. https://doi.org/10.1007/s11060-006-9118-0
Kamarajugadda S, Cai Q, Chen H, Nayak S, Zhu J, He M, et al. Manganese superoxide dismutase promotes anoikis resistance and tumor metastasis. Cell Death Dis. 2013;4:e504. https://doi.org/10.1038/cddis.2013.20
Yeung BHY, Wong KY, Lin MC, Wong CKC, Mashima T, Tsuruo T, et al. Chemosensitisation by manganese superoxide dismutase inhibition is caspase-9 dependent and involves extracellular signal-regulated kinase I/2. Br J Cancer. 2008;99(2):283–93. https://doi.org/10.1038/sj.bjc.6604477
Hu H, Luo LM, Du XL, Feng YB, Zhang Y, Shen XM, et al. Up-regulated manganese superoxide dismutase expression increases apoptosis resistance in human esophageal squamous cell carcinomas. Chin Med J. 2007;120(23):2092–8.
Hardiany NS, Mulyawan W, Wanandi SI. Correlation between oxidative stress and tumor grade in glioma cells from patients in Jakarta. Med J Indones. 2012;21(3):122–7. https://doi.org/10.13181/mji.v21i3.492
Comhair SA, Xu W, Ghosh S, Thunnissen F, Almasan A, Calhoun WJ, et al. Superoxide dismutase inactivation in pathophysiology of asthamatic airway remodeling and reactivity. Am J Pathol. 2005;166(3):663–74. https://doi.org/10.1016/S0002-9440(10)62288-2
Hardiany NS, Sadikin M, Wanandi SI. Gene expression of manganese superoxide dismutase (MnSOD) in human glioma cells. Med J Indones. 2010;19(1):21–5. https://doi.org/10.13181/mji.v19i1.379
Mao W, Chen X, Yang T, Yin Y, Ge M, Luo M, et al. A rapid fluorescent screening method for cellular sensitivity to anti-cancer compound. Cytotechnology. 2012;64(4):451–7. https://doi.org/10.1007/s10616-011-9423-0
Viola G, Salvador A, Vedaldi D, Fortunato E, Disaro S, Basso G, et al. Induction of apoptosis by photoexcited tetracyclic compounds derivatives of benzo[b]thiophenes and pyridines. J Photochemist Photobiol B: Biol. 2006;82(2):105–16. https://doi.org/10.1016/j.jphotobiol.2005.09.004
Hempel N, Carrico PM, Melendez JA. Manganese superoxide dismutase (Sod2) and redox-control of signaling events that drive metastasis. Anticancer Agents Med Chem. 2011;11(2):191–201. https://doi.org/10.2174/187152011795255911
Reddy SP, Britto R, Vinnakota K, Aparna H, Sreepathi HK, Thota B, et al. Novel glioblastoma multiforme markers with diagnostic and prognostic value identified through transcriptome analysis. Clin Cancer Res. 2008;14(10):2978–87. https://doi.org/10.1158/1078-0432.CCR-07-4821
Kalyanaraman B, Darley-Usmar V, Davies KJ, Dennery PA, Forman HJ, Grisham MB. Measuring reactive oxygen and nitrogen species with fluorescent probes: challenges and limitations. Free Radic Biol Med. 2012;52(1):1–6. https://doi.org/10.1016/j.freeradbiomed.2011.09.030
Teoh ML, Sun W, Smith BJ, Oberley LW, Cullen JJ. Modulation of reactive oxygen species in pancreatic cancer. Clin Cancer Res. 2007;13(24):7441–50. https://doi.org/10.1158/1078-0432.CCR-07-0851
Kuninaka S, Ichinose Y, Koja K, Toh Y. Suppression of manganese superoxide dismutase augments sensitivity to radiation, hyperthermia and doxorubicin in colon cancer cell lines by inducing apoptosis. Br J Cancer. 2000;83(7):928–34. https://doi.org/10.1054/bjoc.2000.1367
Hileman EA, Achanta G, Huang P. Superoxide dismutase: an emerging target for cancer therapeutics. Expert Opin Ther Targets. 2001;5(6):697–710. https://doi.org/10.1517/1472822.214.171.1247
Murphy BM, Seamus JM. Caspase structure, activation pathways and substrate. In: Yin XM, Dong Z, editors. Essentials of apoptosis. New Jersey: Humana Press, 2003; p. 3–9. https://doi.org/10.1385/1-59259-361-5:3
Gerhard Kauss. Biochemistry of signal transduction and regulation. 2nd ed. Wiley-VCH 2001; p.351–6.
Dunning S, Hannivoort RA, de Boer JF, Buist-Homan M, Faber KN, Moshage H. Superoxide anions and hydrogen peroxide inhibit proliferation of activated rat stellate cells and induce different modes of cell death. Liver Int. 2009;29(6):922–32. https://doi.org/10.1111/j.1478-3231.2009.02004.x
Zanetti M, Zwacka R, Engelhardt J, Katusic Z, O'Brien T. Superoxide anion and endothelial cell proliferation in normoglycemia and hyperglycemia. Arterioscler Thromb Vasc Biol. 2001;21(2):195–200. https://doi.org/10.1161/01.ATV.21.2.195
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