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


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


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.


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, Dohadw