PX-12 inhibits the growth of A549 lung cancer cells via G2/M phase arrest and ROS-dependent apoptosis

You,Bo  Ra; Shin,Hye  Rim; Park,Woo  Hyun

doi:10.3892/ijo.2013.2152

PX-12 inhibits the growth of A549 lung cancer cells via G2/M phase arrest and ROS-dependent apoptosis

Authors:
- Bo Ra You
- Hye Rim Shin
- Woo Hyun Park
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Affiliations: Department of Physiology, Medical School, Research Institute for Endocrine Sciences, Chonbuk National University, Jeonju 561-180, Republic of Korea
Published online on: October 29, 2013 https://doi.org/10.3892/ijo.2013.2152
Pages: 301-308

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Abstract

PX-12 (1-methylpropyl 2-imidazolyl disulfide) is an inhibitor of thioredoxin (Trx-1), which has antitumor effects. However, little is known about the toxicological effect of PX-12 on cancer cells. We investigated the anti-growth effects of PX-12 on A549 lung cancer cells in relation to reactive oxygen species (ROS) and glutathione (GSH) levels. Based on MTT assays, PX-12 inhibited the growth of A549 cells with an IC50 of approximately 20 µM at 72 h. DNA flow cytometric analysis indicated that PX-12 significantly induced the G2/M phase arrest of the cell cycle in A549 cells. This agent also induced apoptotic cell death, as demonstrated by Annexin V-FITC staining cells and the loss of mitochondrial membrane potential MMP (∆ψm). In addition, the administration of Bax siRNA attenuated PX-12-induced A549 cell death. All the tested caspase inhibitors, especially Z-VAD significantly prevented apoptosis induced by PX-12. With respect to ROS and GSH levels, PX-12 increased ROS levels including O2•- in A549 cells and induced GSH depletion. N-acetyl cysteine (NAC) markedly reduced ROS levels in PX-12-treated A549 cells. NAC also prevented apoptotic cell death and GSH depletion induced by PX-12. This is the first report to show that PX-12 inhibits the growth of A549 cells via G2/M phase arrest, and Bax-mediated and ROS-dependent apoptosis.

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1.	Li C, Thompson MA, Tamayo AT, et al: Over-expression of Thioredoxin-1 mediates growth, survival, and chemoresistance and is a druggable target in diffuse large B-cell lymphoma. Oncotarget. 3:314–326. 2012.PubMed/NCBI
2.	Yang J, Li C, Ding L, Guo Q, You Q and Jin S: Gambogic acid deactivates cytosolic and mitochondrial thioredoxins by covalent binding to the functional domain. J Nat Prod. 75:1108–1116. 2012. View Article : Google Scholar : PubMed/NCBI
3.	Chae JS, Gil Hwang S, Lim DS and Choi EJ: Thioredoxin-1 functions as a molecular switch regulating the oxidative stress-induced activation of MST1. Free Radic Biol Med. 53:2335–2343. 2012. View Article : Google Scholar : PubMed/NCBI
4.	Ungerstedt J, Du Y, Zhang H, Nair D and Holmgren A: In vivo redox state of human thioredoxin and redox shift by the histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA). Free Radic Biol Med. 53:2002–2007. 2012. View Article : Google Scholar : PubMed/NCBI
5.	Lim JY, Yoon SO, Hong SW, Kim JW, Choi SH and Cho JY: Thioredoxin and thioredoxin-interacting protein as prognostic markers for gastric cancer recurrence. World J Gastroenterol. 18:5581–5588. 2012. View Article : Google Scholar : PubMed/NCBI
6.	Pramanik KC and Srivastava SK: Apoptosis signal-regulating kinase 1-thioredoxin complex dissociation by capsaicin causes pancreatic tumor growth suppression by inducing apoptosis. Antioxid Redox Signal. 17:1417–1432. 2012. View Article : Google Scholar
7.	Dunn LL, Buckle AM, Cooke JP and Ng MK: The emerging role of the thioredoxin system in angiogenesis. Arterioscler Thromb Vasc Biol. 30:2089–2098. 2010. View Article : Google Scholar : PubMed/NCBI
8.	Cha MK, Suh KH and Kim IH: Overexpression of peroxiredoxin I and thioredoxin 1 in human breast carcinoma. J Exp Clin Cancer Res. 28:932009. View Article : Google Scholar : PubMed/NCBI
9.	Wondrak GT: Redox-directed cancer therapeutics: molecular mechanisms and opportunities. Antioxid Redox Signal. 11:3013–3069. 2009. View Article : Google Scholar : PubMed/NCBI
10.	Welsh SJ, Williams RR, Birmingham A, Newman DJ, Kirkpatrick DL and Powis G: The thioredoxin redox inhibitors 1-methylpropyl 2-imidazolyl disulfide and pleurotin inhibit hypoxia-induced factor 1alpha and vascular endothelial growth factor formation. Mol Cancer Ther. 2:235–243. 2003.
11.	Mukherjee A and Martin SG: The thioredoxin system: a key target in tumour and endothelial cells. Br J Radiol. 81(Spec1): S57–S68. 2008. View Article : Google Scholar : PubMed/NCBI
12.	Baker AF, Adab KN, Raghunand N, et al: A phase IB trial of 24-hour intravenous PX-12, a thioredoxin-1 inhibitor, in patients with advanced gastrointestinal cancers. Invest New Drugs. 31:631–641. 2013. View Article : Google Scholar : PubMed/NCBI
13.	Ramanathan RK, Kirkpatrick DL, Belani CP, et al: A Phase I pharmacokinetic and pharmacodynamic study of PX-12, a novel inhibitor of thioredoxin-1, in patients with advanced solid tumors. Clin Cancer Res. 13:2109–2114. 2007. View Article : Google Scholar : PubMed/NCBI
14.	Petty RD, Nicolson MC, Kerr KM, Collie-Duguid E and Murray GI: Gene expression profiling in non-small cell lung cancer: from molecular mechanisms to clinical application. Clin Cancer Res. 10:3237–3248. 2004. View Article : Google Scholar : PubMed/NCBI
15.	Fernandes AP, Capitanio A, Selenius M, Brodin O, Rundlof AK and Bjornstedt M: Expression profiles of thioredoxin family proteins in human lung cancer tissue: correlation with proliferation and differentiation. Histopathology. 55:313–320. 2009. View Article : Google Scholar : PubMed/NCBI
16.	Wangpaichitr M, Sullivan EJ, Theodoropoulos G, et al: The relationship of thioredoxin-1 and cisplatin resistance: its impact on ROS and oxidative metabolism in lung cancer cells. Mol Cancer Ther. 11:604–615. 2012. View Article : Google Scholar : PubMed/NCBI
17.	Han YH, Kim SZ, Kim SH and Park WH: Pyrogallol inhibits the growth of lung cancer Calu-6 cells via caspase-dependent apoptosis. Chem Biol Interact. 177:107–114. 2009. View Article : Google Scholar : PubMed/NCBI
18.	Han YH and Park WH: The effects of N-acetyl cysteine, buthionine sulfoximine, diethyldithiocarbamate or 3-amino-1,2,4-triazole on antimycin A-treated Calu-6 lung cells in relation to cell growth, reactive oxygen species and glutathione. Oncol Rep. 22:385–391. 2009.
19.	Han YH, Moon HJ, You BR, Kim SZ, Kim SH and Park WH: Effects of carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone on the growth inhibition in human pulmonary adenocarcinoma Calu-6 cells. Toxicology. 265:101–107. 2009. View Article : Google Scholar : PubMed/NCBI
20.	You BR and Park WH: Zebularine inhibits the growth of HeLa cervical cancer cells via cell cycle arrest and caspase-dependent apoptosis. Mol Biol Rep. 39:9723–9731. 2012. View Article : Google Scholar : PubMed/NCBI
21.	Han YH, Moon HJ, You BR and Park WH: The effect of MG132, a proteasome inhibitor on HeLa cells in relation to cell growth, reactive oxygen species and GSH. Oncol Rep. 22:215–221. 2009.PubMed/NCBI
22.	Han YH, Kim SH, Kim SZ and Park WH: Carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP) as an O₂(^*-) generator induces apoptosis via the depletion of intracellular GSH contents in Calu-6 cells. Lung Cancer. 63:201–209. 2009. View Article : Google Scholar : PubMed/NCBI
23.	Han YH and Park WH: Propyl gallate inhibits the growth of HeLa cells via regulating intracellular GSH level. Food Chem Toxicol. 47:2531–2538. 2009. View Article : Google Scholar : PubMed/NCBI
24.	You BR and Park WH: Gallic acid-induced lung cancer cell death is related to glutathione depletion as well as reactive oxygen species increase. Toxicol In Vitro. 24:1356–1362. 2010. View Article : Google Scholar : PubMed/NCBI
25.	Griffiths EJ: Mitochondria - potential role in cell life and death. Cardiovasc Res. 46:24–27. 2000. View Article : Google Scholar : PubMed/NCBI
26.	Vogt A, Tamura K, Watson S and Lazo JS: Antitumor imidazolyl disulfide IV-2 causes irreversible G(2)/M cell cycle arrest without hyperphosphorylation of cyclin-dependent kinase Cdk1. J Pharmacol Exp Ther. 294:1070–1075. 2000.PubMed/NCBI
27.	Yang J, Liu X, Bhalla K, et al: Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science. 275:1129–1132. 1997. View Article : Google Scholar : PubMed/NCBI
28.	Martinou JC and Youle RJ: Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics. Dev Cell. 21:92–101. 2011. View Article : Google Scholar : PubMed/NCBI
29.	Lee YJ, Kim JH, Chen J and Song JJ: Enhancement of metabolic oxidative stress-induced cytotoxicity by the thioredoxin inhibitor 1-methylpropyl 2-imidazolyl disulfide is mediated through the ASK1-SEK1-JNK1 pathway. Mol Pharmacol. 62:1409–1417. 2002. View Article : Google Scholar
30.	Estrela JM, Ortega A and Obrador E: Glutathione in cancer biology and therapy. Crit Rev Clin Lab Sci. 43:143–181. 2006. View Article : Google Scholar
31.	You BR and Park WH: Arsenic trioxide induces human pulmonary fibroblast cell death via increasing ROS levels and GSH depletion. Oncol Rep. 28:749–757. 2012.PubMed/NCBI