Cyclooxygenase inhibitors combined with deuterium-enriched water augment cytotoxicity in A549 lung cancer cell line via activation of apoptosis and MAPK pathways

Document Type: Original Article

Authors

1 Pharmaceutics Research Centre, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

2 Department of Medicinal Chemistry, School of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran

3 Neuroscience Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

4 Department of Pharmacology & Toxicology, School of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran

5 5 Physiology Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

Abstract

Objective(s): Combination chemotherapy is a rational strategy to increase patient response and tolerability and to decrease adverse effects and drug resistance. Recently, the use of non-steroidal anti-inflammatory drugs (NSAIDs) has been reported to be associated with reduction in occurrence of a variety of cancers including lung cancer. On the other hand, growing evidences suggest that deuterium-enriched water (DEW, D2O) and deuterium-depleted water (DDW) play a role both in treatment and prevention of cancers. In the present study, we examined the effects of DEW and DDW in combination with two NSAIDs, celecoxib and indomethacin, on A549 human non-small lung cancer cell to identify novel treatment options.
Materials and Methods: The cytotoxicity of celecoxib or indomethacin, alone and in combination with DDW and DEW was determined. The COX-2, MAPK pathway proteins, the anti-apoptotic Bcl2 and pro-apoptotic Bax proteins and caspase-3 activity were studied for cytotoxic combinations.
Results: Co-administration of selective and non-selective COX-2 inhibitors with DEW led to a remarkable increase in cytotoxicity and apoptosis of A549 cells. These events were associated with activation of p38 and JNK MAPKs and decreasing pro-survival proteins Bcl-2, COX-2 and ERK1/2. Furthermore, the combination therapy activated caspase-3, and the apoptosis mediator, and disabled poly ADP-ribose polymerase (PARP), the key DNA repair enzyme, by cleaving it.  
Conclusion: The combination of DEW with NSAIDs might be effective against lung cancer cells by influence on principal cell signalling pathways, and this has a potential to become a candidate for chemotherapy.

Keywords

Main Subjects


1. Torre LA, Siegel RL, Jemal A. Lung cancer statistics. In: Ahmad A, Gadgeel, Shirish, editor. Lung Cancer and Personalized Medicine: Springer; 2016. p. 1-19.
2. Reck M, Rodríguez-Abreu D, Robinson AG, Hui R, Csőszi T, Fülöp A, et al. Pembrolizumab versus chemotherapy for PD-L1–positive non–small-cell lung cancer. N Engl J Med 2016; 2016:1823-1833.
3. Solomon BJ, Mok T, Kim DW, Wu YL, Nakagawa K, Mekhail T, et al. First-line crizotinib versus chemotherapy in ALK-positive lung cancer. N Engl J Med 2014; 371:2167-2177.
4. Huang Xz, Chen Y, Wu J, Zhang X, Wu Cc, Zhang Cy, et al. Aspirin and non-steroidal anti-inflammatory drugs use reduce gastric cancer risk: A dose-response meta-analysis. Oncotarget 2017; 8:4781-4795.
5. Cha BK, Kim YS, Hwang KE, Cho KH, Oh SH, Kim BR, et al. Celecoxib and sulindac inhibit TGF-β1-induced epithelial-mesenchymal transition and suppress lung cancer migration and invasion via downregulation of sirtuin 1. Oncotarget 2016; 7:57213-57227.
6. Baik CS, Brasky TM, Pettinger M, Luo J, Gong Z, Wactawski-Wende J, et al. Non-steroidal anti-inflammatory drug and aspirin use in relation to lung cancer risk among postmenopausal women. Cancer Epidemiol Biomarkers Prev 2015; 24:790-797.
7. Valle BL, D’Souza T, Becker KG, Wood III WH, Zhang Y, Wersto RP, et al. Non-steroidal anti-inflammatory drugs decrease E2F1 expression and inhibit cell growth in ovarian cancer cells. PLoS One 2013; 8:e61836.
8. Guadagni F, Ferroni P, Palmirotta R, Del Monte G, Formica V, Roselli M. Non-steroidal anti-inflammatory drugs in cancer prevention and therapy. Anticancer Res 2007; 27:3147-3162.
9. Day RO, Graham GG. Non-steroidal Anti-inflammatory Drugs: Overview. Compendium of Inflammatory Diseases 2016:1-9.
10. Ettarh R, Cullen A, Calamai A. NSAIDs and cell proliferation in colorectal cancer. Pharmaceuticals 2010; 3:2007-2021.
11. Rai N, Sarkar M, Raha S. Piroxicam, a traditional non-steroidal anti-inflammatory drug (NSAID) causes apoptosis by ROS mediated Akt activation. Pharmacol Rep 2015; 67:1215-1223.
12. Chen L, He Y, Huang H, Liao H, Wei W. Selective COX-2 inhibitor celecoxib combined with EGFR-TKI ZD1839 on non-small cell lung cancer cell lines: in vitro toxicity and mechanism study. Med Oncol 2008; 25:161-171.
13. Wang Zl, Fan Zq, Jiang Hd, Qu Jm. Selective Cox-2 inhibitor celecoxib induces epithelial-mesenchymal transition in human lung cancer cells via activating MEK-ERK signaling. Carcinogenesis 2012; 34:638-646.
14. Krysan K, Reckamp KL, Dalwadi H, Sharma S, Rozengurt E, Dohadwala M, et al. Prostaglandin E2 activates mitogen-activated protein kinase/Erk pathway signaling and cell proliferation in non-small cell lung cancer cells in an epidermal growth factor receptor-independent manner. Cancer Res 2005; 65:6275-6281.
15. Mandegary A, Torshabi M, Seyedabadi M, Amirheidari B, Sharif E, Ghahremani MH. Indomethacin-enhanced anticancer effect of arsenic trioxide in A549 cell line: involvement of apoptosis and phospho-ERK and p38 MAPK pathways. Biomed Res Int 2013; 2013:237543.
16. Jin HO, Seo SK, Woo SH, Lee HC, Kim ES, Yoo DH, et al. A combination of sulindac and arsenic trioxide synergistically induces apoptosis in human lung cancer H1299 cells via c-Jun NH2-terminal kinase-dependent Bcl-xL phosphorylation. Lung Cancer 2008; 61:317-327.
17. Han YH, Kim SZ, Kim SH, Park WH. Induction of apoptosis in arsenic trioxide-treated lung cancer A549 cells by buthionine sulfoximine. Mol Cells 2008; 26:158-164.
18. Schroeder CP, Kadara H, Lotan D, Woo JK, Lee HY, Hong WK, et al. Involvement of mitochondrial and Akt signaling pathways in augmented apoptosis induced by a combination of low doses of celecoxib and N-(4-hydroxyphenyl) retinamide in premalignant human bronchial epithelial cells. Cancer Res 2006; 66:9762-9770.
19. Gyöngyi Z, Budán F, Szabó I, Ember I, Kiss I, Krempels K, et al. Deuterium depleted water effects on survival of lung cancer patients and expression of Kras, Bcl2, and Myc genes in mouse lung. Nutr Cancer 2013; 65:240-246.
20. Gyongyi Z, Somlyai G. Deuterium depletion can decrease the expression of c-myc, Ha-ras and p53 gene in carcinogen-treated mice. In vivo 2000; 14:437-440.
21. Kovács A, Guller I, Krempels K, Somlyai I, Jánosi I, Gyomgyi Z, et al. Deuterium depletion may delay the progression of prostate cancer. J Cancer Ther 2011; 2:548.
22. Wang H, Zhu B, He Z, Fu H, Dai Z, Huang G, et al. Deuterium-depleted water (DDW) inhibits the proliferation and migration of nasopharyngeal carcinoma cells in vitro. Biomed Pharmacother 2013; 67:489-496.
23. Mirică RE. Deuterium-depleted water in cancer therapy. Environmental Engineering & Management Journal (EEMJ) 2010; 9.
24. COng FS, ZhanG Yr, SHeng HC, Ao ZH, ZhanG SY, WAng Jy. Deuterium-depleted water inhibits human lung carcinoma cell growth by apoptosis. Exp Ther Med 2010; 1:277-283.
25. Altermatt HJ, Gebbers JO, Laissue JA. Heavy water delays growth of human carcinoma in nude mice. Cancer 1988; 62:462-466.
26. Hatta J, Hatta T, Moritake K, Otani H. Heavy water inhibiting the expression of transforming growth factor–β1 and the development of kaolin-induced hydrocephalus in mice. J Neurosurg: Pediatrics 2006; 104:251-258.
27. Soleyman-Jahi S, Zendehdel K, Akbarzadeh K, Haddadi M, Amanpour S, Muhammadnejad S. In vitro assessment of antineoplastic effects of deuterium depleted water. Asian Pac J Cancer Prev 2014; 15:2179-2183.
28. Takeda H, Nio Y, Omori H, Uegaki K, Hirahara N, Sasaki S, et al. Mechanisms of cytotoxic effects of heavy water (deuterium oxide: D20) on cancer cells. Anticancer Drug 1998; 9:715-725.
29. Kushner D, Baker A, Dunstall T. Pharmacological uses and perspectives of heavy water and deuterated compounds. Can J Physiol Pharmacol 1999; 77:79-88.
30. Krempels K, Somlyai I, Somlyai G. A retrospective evaluation of the effects of deuterium depleted water consumption on 4 patients with brain metastases from lung cancer. Integr Cancer Ther 2008; 7:172-181.
31. Cong FS, Zhang YR, Sheng HC, Ao ZH, Zhang SY, Wang JY. Deuterium-depleted water inhibits human lung carcinoma cell growth by apoptosis. Exp Ther Med 2010; 1:277-283.
32. Bader Y, Hartmann J, Horvath Z, Saiko P, Grusch M, Madlener S, et al. Synergistic effects of deuterium oxide and gemcitabine in human pancreatic cancer cell lines. Cancer Lett 2008; 259:231-239.
33. Freshney RI. culture of animal cells: Oxford University Press; 1992.
34. Friedrich M, Reichert K, Woeste A, Polack S, Fischer D, Hoellen F, et al. Effects of Combined Treatment with Vitamin D and COX2 Inhibitors on Breast Cancer Cell Lines. Anticancer Res 2018; 38:1201-1207.
35. Bayat Mokhtari R, Homayouni TS, Baluch N, Morgatskaya E, Kumar S, Das B, et al. Combination therapy in combating cancer. Oncotarget 2017; 8:38022-38043.
36. Mandegary A, Mehrabani M. Effects of arsenic trioxide, all-trans-retinoic acid and dexamethasone on NB4 cell line. Daru 2010; 18:303-309.
37. Iwama E, Nakanishi Y, Okamoto I. Combined therapy with epidermal growth factor receptor tyrosine kinase inhibitors for non-small cell lung cancer. Expert Rev Anticancer Ther 2018:1-10.
38. Gyongyi Z, Budan F, Szabo I, Ember I, Kiss I, Krempels K, et al. Deuterium depleted water effects on survival of lung cancer patients and expression of Kras, Bcl2, and Myc genes in mouse lung. Nutr Cancer 2013; 65:240-246.
39. Hartmann J, Bader Y, Horvath Z, Saiko P, Grusch M, Illmer C, et al. Effects of heavy water (D2O) on human pancreatic tumor cells. Anticancer Res 2005; 25:3407-3411.
40. Bahk JY, Lee JH, Chung HS, Lee HY, Chung BC, Park MS, et al. Anticancer effect of deuterium oxide on a bladder cancer cell related to Bcl-2 and Bax.  2007; 13:501-507.
41. Noguera Aguilar JF, Amengual Antich I, Pujol Tugores JJ. Dose of rofecoxib in colorectal cancer. Int J Cancer 2004; 110:309; author reply 310.
42. Patel MI, Subbaramaiah K, Du B, Chang M, Yang P, Newman RA, et al. Celecoxib inhibits prostate cancer growth: evidence of a cyclooxygenase-2-independent mechanism. Clin Cancer Res 2005; 11:1999-2007.
43. Abou-Issa H, Alshafie G. Celecoxib: a novel treatment for lung cancer. Expert Rev Anticancer Ther 2004; 4:725-734.
44. Park JH, Kim EJ, Jang HY, Shim H, Lee KK, Jo HJ, et al. Combination treatment with arsenic trioxide and sulindac enhances apoptotic cell death in lung cancer cells via activation of oxidative stress and mitogen-activated protein kinases. Oncol Rep 2008; 20:379-384.
45. Guo YC, Chang CM, Hsu WL, Chiu SJ, Tsai YT, Chou YH, et al. Indomethacin inhibits cancer cell migration via attenuation of cellular calcium mobilization. Molecules 2013; 18:6584-6596.
46. Groen HJ, Sietsma H, Vincent A, Hochstenbag MM, van Putten JW, van den Berg A, et al. Randomized, placebo-controlled phase III study of docetaxel plus carboplatin with celecoxib and cyclooxygenase-2 expression as a biomarker for patients with advanced non-small-cell lung cancer: the NVALT-4 study. J Clin Oncol 2011; 29:4320-4326.
47. Roca-Ferrer J, Pujols L, Agusti C, Xaubet A, Mullol J, Gimferrer JM, et al. [Cyclooxigenase-2 levels are increased in the lung tissue and bronchial tumors of patients with chronic obstructive pulmonary disease]. Arch Bronconeumol 2011; 47:584-589.
48. Soslow RA, Dannenberg AJ, Rush D, Woerner BM, Khan KN, Masferrer J, et al. COX-2 is expressed in human pulmonary, colonic, and mammary tumors. Cancer 2000; 89:2637-2645.
49. Li F, Liu Y, Chen H, Liao D, Shen Y, Xu F, et al. EGFR and COX-2 protein expression in non-small cell lung cancer and the correlation with clinical features. J Exp Clin Cancer Res 2011; 30:27.
50. Greenhough A, Smartt HJ, Moore AE, Roberts HR, Williams AC, Paraskeva C, et al. The COX-2/PGE2 pathway: key roles in the hallmarks of cancer and adaptation to the tumour microenvironment. Carcinogenesis 2009; 30:377-386.
51. Xu L, Stevens J, Hilton MB, Seaman S, Conrads TP, Veenstra TD, et al. COX-2 inhibition potentiates antiangiogenic cancer therapy and prevents metastasis in preclinical models. Sci Transl Med 2014; 6:242ra284.
52. Wang MT, Honn KV, Nie D. Cyclooxygenases, prostanoids, and tumor progression. Cancer Metastasis Rev 2007; 26:525-534.
53. Yoshinaka R, Shibata MA, Morimoto J, Tanigawa N, Otsuki Y. COX-2 inhibitor celecoxib suppresses tumor growth and lung metastasis of a murine mammary cancer. Anticancer Res 2006; 26:4245-4254.
54. Cusimano A, Fodera D, D’Alessandro N, Lampiasi N, Azzolina A, Montalto G, et al. Potentiation of the antitumor effects of both selective cyclooxygenase-1 and cyclooxygenase-2 inhibitors in human hepatic cancer cells by inhibition of the MEK/ERK pathway. Cancer Biol Ther 2007; 6:1461-1468.
55. Tegeder I, Pfeilschifter J, Geisslinger G. Cyclooxygenase-independent actions of cyclooxygenase inhibitors. FASEB J 2001; 15:2057-2072.
56. Han S, Roman J. COX-2 inhibitors suppress lung cancer cell growth by inducing p21 via COX-2 independent signals. Lung Cancer 2006; 51:283-296.
57. Sun Y, Sinicrope FA. Selective inhibitors of MEK1/ERK44/42 and p38 mitogen-activated protein kinases potentiate apoptosis induction by sulindac sulfide in human colon carcinoma cells. Mol Cancer Ther 2005; 4:51-59.
58. Gao J, Niwa K, Takemura M, Sun W, Onogi K, Wu Y, et al. Significant anti-proliferation of human endometrial cancer cells by combined treatment with a selective COX-2 inhibitor NS398 and specific MEK inhibitor U0126. Int J Oncol 2005; 26:737-744.
59. Rice PL, Beard KS, Driggers LJ, Ahnen DJ. Inhibition of extracellular-signal regulated kinases 1/2 is required for apoptosis of human colon cancer cells in vitro by sulindac metabolites. Cancer Res 2004; 64:8148-8151.
60. Husain SS, Szabo IL, Pai R, Soreghan B, Jones MK, Tarnawski AS. MAPK (ERK2) kinase--a key target for NSAIDs-induced inhibition of gastric cancer cell proliferation and growth. Life Sci 2001; 69:3045-3054.
61. Uemura T, Moritake K, Akiyama Y, Kimura Y, Shingu T, Yamasaki T. Experimental validation of deuterium oxide-mediated antitumoral activity as it relates to apoptosis in murine malignant astrocytoma cells. J Neurosurg 2002; 96:900-908.
62. Liggett JL, Min KW, Smolensky D, Baek SJ. A novel COX-independent mechanism of sulindac sulfide involves cleavage of epithelial cell adhesion molecule protein. Exp Cell Res 2014; 326:1-9.
63. Dhillon AS, Hagan S, Rath O, Kolch W. MAP kinase signalling pathways in cancer. Oncogene 2007; 26:3279-3290.
64. Huang D, Ichikawa K. Drug discovery targeting


apoptosis. Nat Rev Drug Discov 2008; 7:1041.
65. Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 1995; 270:1326-1331.
66. Wang X, Martindale JL, Holbrook NJ. Requirement for ERK activation in cisplatin-induced apoptosis. J Biol Chem 2000; 275:39435-39443.
67. Poligone B, Baldwin AS. Positive and negative regulation of NF-kappaB by COX-2: roles of different prostaglandins. J Biol Chem 2001; 276:38658-38664.
68. Porebska I, Wyrodek E, Kosacka M, Adamiak J, Jankowska R, Harlozinska-Szmyrka A. Apoptotic markers p53, Bcl-2 and Bax in primary lung cancer. In Vivo 2006; 20:599-604.
69. Jendrossek V, Handrick R, Belka C. Celecoxib activates a novel mitochondrial apoptosis signaling pathway. FASEB J 2003; 17:1547-1549.