Effects of ERK1/2 S-nitrosylation on ERK1/2 phosphorylation and cell survival in glioma cells

Jin,Lei; Cao,Yujia; Zhang,Tong; Wang,Peng; Ji,Daofei; Liu,Xuejiao; Shi,Hengliang; Hua,Lei; Yu,Rutong; Gao,Shangfeng

doi:10.3892/ijmm.2017.3334

Effects of ERK1/2 S-nitrosylation on ERK1/2 phosphorylation and cell survival in glioma cells

Authors:
- Lei Jin
- Yujia Cao
- Tong Zhang
- Peng Wang
- Daofei Ji
- Xuejiao Liu
- Hengliang Shi
- Lei Hua
- Rutong Yu
- Shangfeng Gao
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Affiliations: Institute of Nervous System Diseases, Xuzhou Medical University, Xuzhou, Jiangsu 221002, P.R. China, Department of Neurosurgery, People's Hospital of Gaoxin District, Suzhou, Jiangsu 215011, P.R. China, Department of Neurosurgery, Brain Hospital, Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221002, P.R. China
Published online on: December 20, 2017 https://doi.org/10.3892/ijmm.2017.3334
Pages: 1339-1348
Copyright: © Jin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Aberrant activation of extracellular signal‑regulated kinase 1/2 (ERK1/2) by phosphorylation modification can trigger tumor cell development in glioma. S‑nitrosylation, which refers to the covalent addition of a nitric oxide (NO) group to a cysteine (Cys) thiol, is an important post‑translational modification that occurs on numerous cancer‑associated proteins. Protein S‑nitrosylation can increase or decrease protein activity and stability, and subsequent signal transduction and cellular processes. However, the association between ERK1/2 S‑nitrosylation and ERK1/2 phosphorylation, and the effects of ERK1 S‑nitrosylation on glioma cell survival are currently unknown. U251 glioma cells were treated with NO donors sodium nitroprusside (SNP) or S‑nitrosoglutathione (GSNO). CCK8 assay was used to assess the cell viability. NO levels in the medium were detected by Griess assay. Western blot analysis and biotin switch assay were employed to detect the ERK1/2 phosphorylation and S-nitrosylation. ERK1 wild-type and mutant plasmids were constructed, and used to transfect the U251 cells. Caspase-3 western blot analysis and flow cytometry were employed to assess cell apoptosis. The present study demonstrated that treatment with the NO donors SNP or GSNO led to an increase in ERK1/2 S‑nitrosylation, and a reduction in ERK1/2 phosphorylation, which was accompanied by growth inhibition of U251 glioma cells. Mutational analysis demonstrated that Cys183 was vital for S‑nitrosylation of ERK1, and that preventing ERK1 S‑nitrosylation by replacing Cys183 with alanine partially reversed GSNO‑induced cell apoptosis, and reductions in cell viability and ERK1/2 phosphorylation. In addition, increased ERK1/2 phosphorylation was associated with decreased ERK1/2 S‑nitrosylation in human glioma tissues. These findings identified the relationship between ERK1/2 S‑nitrosylation and phosphorylation in vitro and in vivo, and revealed a novel mechanism of ERK1/2 underlying tumor cell development and apoptotic resistance of glioma.

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1	Stupp R, Hegi ME, Gilbert MR and Chakravarti A: Chemoradiotherapy in malignant glioma: Standard of care and future directions. J Clin Oncol. 25:4127–4136. 2007. View Article : Google Scholar : PubMed/NCBI
2	Sanai N, Alvarez-Buylla A and Berger MS: Neural stem cells and the origin of gliomas. N Engl J Med. 353:811–822. 2005. View Article : Google Scholar : PubMed/NCBI
3	Wen PY and Kesari S: Malignant gliomas in adults. N Engl J Med. 359:492–507. 2008. View Article : Google Scholar : PubMed/NCBI
4	Roberts PJ and Der CJ: Targeting the Raf-MEK-ERK mitogen-activated protein kinase cascade for the treatment of cancer. Oncogene. 26:3291–3310. 2007. View Article : Google Scholar : PubMed/NCBI
5	Pandey V, Bhaskara VK and Babu PP: Implications of mitogen-activated protein kinase signaling in glioma. J Neurosci Res. 94:114–127. 2016. View Article : Google Scholar
6	Dong Chen, Waters SB, Holt KH and Pessin JE: SOS phosphorylation and disassociation of the Grb2-SOS complex by the ERK and JNK signaling pathways. J Biol Chem. 271:6328–6332. 1996. View Article : Google Scholar
7	Bhaskara VK, Panigrahi M, Challa S and Babu PP: Comparative status of activated ERK1/2 and PARP cleavage in human gliomas. Neuropathology. 25:48–53. 2005. View Article : Google Scholar : PubMed/NCBI
8	Xie H, Xue YX, Liu LB, Wang P, liu YH and Ying HQ: Expressions of matrix metalloproteinase-7 and matrix metalloproteinase-14 associated with the activation of extracellular signal-regulated kinase1/2 in human brain gliomas of different pathological grades. Med Oncol. 28(Suppl 1): S433–S438. 2011. View Article : Google Scholar
9	Lundberg JO, Gladwin MT and Weitzberg E: Strategies to increase nitric oxide signalling in cardiovascular disease. Nat Rev Drug Discov. 14:623–641. 2015. View Article : Google Scholar : PubMed/NCBI
10	Nakamura T and Lipton SA: Protein S-nitrosylation as a therapeutic target for neurodegenerative diseases. Trends Pharmacol Sci. 37:73–84. 2016. View Article : Google Scholar :
11	Wang Z: Protein S-nitrosylation and cancer. Cancer Lett. 320:123–129. 2012. View Article : Google Scholar : PubMed/NCBI
12	Feng X, Sun T, Bei Y, Ding S, Zheng W, lu Y and Shen P: S-nitrosylation of ERK inhibits ERK phosphorylation and induces apoptosis. Sci Rep. 3:18142013. View Article : Google Scholar : PubMed/NCBI
13	Shen A, Gao S, Ben Z, Wang H, Jia J, Tao T, Niu S, Li X and Cheng C: Identification and potential role of PSD-95 in Schwann cells. Neurol Sci. 29:321–330. 2008. View Article : Google Scholar : PubMed/NCBI
14	Louis DN, Ohgaki H, Wiestler OD and Cavenee WK: WHO Classification of Tumours of the Central Nervous System. IARC WHO Classification of Tumours; Lyon: 2016
15	Kurimoto M, Endo S, Hirashima Y, Hamada H, Ogiichi T and Takaku A: Growth inhibition and radiosensitization of cultured glioma cells by nitric oxide generating agents. J Neurooncol. 42:35–44. 1999. View Article : Google Scholar : PubMed/NCBI
16	Stamler JS, Toone EJ, Lipton SA and Sucher NJ: (S)NO signals: Translocation, regulation, and a consensus motif. Neuron. 18:691–696. 1997. View Article : Google Scholar : PubMed/NCBI
17	Weyerbrock A, Baumer B and Papazoglou A: Growth inhibition and chemosensitization of exogenous nitric oxide released from NONOates in glioma cells in vitro. J Neurosurg. 110:128–136. 2009. View Article : Google Scholar
18	Maejima Y, Adachi S, Morikawa K, Ito H and Isobe M: Nitric oxide inhibits myocardial apoptosis by preventing caspase-3 activity via S-nitrosylation. J Mol Cell Cardiol. 38:163–174. 2005. View Article : Google Scholar
19	Lechner M, Lirk P and Rieder J: Inducible nitric oxide synthase (iNOS) in tumor biology: The two sides of the same coin. Semin Cancer Biol. 15:277–289. 2005. View Article : Google Scholar : PubMed/NCBI
20	Lancaster JR Jr and Xie K: Tumors face NO problems. Cancer Res. 66:6459–6462. 2006. View Article : Google Scholar : PubMed/NCBI
21	Wang Y, Chen C, Loake GJ and Chu C: Nitric oxide: Promoter or suppressor of programmed cell death. Protein Cell. 1:133–142. 2010. View Article : Google Scholar
22	Azad N, Vallyathan V, Wang L, Tantishaiyakul V, Stehlik C, leonard SS and Rojanasakul Y: S-nitrosylation of Bcl-2 inhibits its ubiquitin-proteasomal degradation. A novel antiapoptotic mechanism that suppresses apoptosis. J Biol Chem. 281:34124–34134. 2006. View Article : Google Scholar : PubMed/NCBI
23	Chanvorachote P, Nimmannit U, Stehlik C, Wang L, Jiang BH, Ongpipatanakul B and Rojanasakul Y: Nitric oxide regulates cell sensitivity to cisplatin-induced apoptosis through S-nitrosylation and inhibition of Bcl-2 ubiquitination. Cancer Res. 66:6353–6360. 2006. View Article : Google Scholar : PubMed/NCBI
24	Leon-Bollotte L, Subramaniam S, Cauvard O, Plenchette-Colas S, Paul C, Godard C, Martinez-Ruiz A, Legembre P, Jeannin JF and Bettaieb A: S-nitrosylation of the death receptor fas promotes fas ligand-mediated apoptosis in cancer cells. Gastroenterology. 140:2009–2018. 2018.e2001–2004. 2011. View Article : Google Scholar : PubMed/NCBI
25	Sebolt-Leopold JS and Herrera R: Targeting the mitogen-activated protein kinase cascade to treat cancer. Nat Rev Cancer. 4:937–947. 2004. View Article : Google Scholar : PubMed/NCBI
26	Murillo-Carretero M, Torroglosa A, Castro C, Villalobo A and Estrada C: S-nitrosylation of the epidermal growth factor receptor: A regulatory mechanism of receptor tyrosine kinase activity. Free Radic Biol Med. 46:471–479. 2009. View Article : Google Scholar
27	Kaliyaperumal K, Sharma A K, McDonald DG, Dhindsa JS, Yount C, Singh AK, Won JS and Singh I: S-nitrosoglutathione-mediated STAT3 regulation in efficacy of radiotherapy and cisplatin therapy in head and neck squamous cell carcinoma. Redox Biol. 6:41–50. 2015. View Article : Google Scholar : PubMed/NCBI
28	Chattopadhyay M, Goswami S, Rodes DB, Kodela R, Velazquez CA, Boring D, Crowell JA and Kashfi K: NO-releasing NSAIDs suppress NF-κB signaling in vitro and in vivo through S-nitrosylation. Cancer Lett. 298:204–211. 2010. View Article : Google Scholar : PubMed/NCBI
29	Szabo C: Gasotransmitters in cancer: From pathophysiology to experimental therapy. Nat Rev Drug Discov. 15:185–203. 2016. View Article : Google Scholar