Evaluation of cytotoxic, membrane, and DNA damaging effects of Thymus revolutus Célak essential oil on different cancer cells

Background/aim: In this study, we evaluated Thymus revolutus Célak essential oil and its two main constituents, γ-terpinene and p-cymene, as potential oxidative agents against lung cancer and epidermoid carcinoma cells. Materials and methods: Cell viability assessment was made by CellTiter-Blue® cell viability and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay after treatment with 5 600 µg/mL concentrations of essential oil, γ-terpinene, and p-cymene. Malondialdehyde and 8-hydroxy-2′-deoxyguanosine levels in parental H1299, epirubicin-resistant (drug-resistant) H1299, A549, and A431 cells were also assayed after essential oil, γ-terpinene, and p-cymene had been administered for 24 h. Glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GRx), and glucose 6-phosphate dehydrogenase (G6PD) activities were determined. Results: Parental H1299 cells were found to be more sensitive to the cytotoxic effects of all compounds. While A431 cells had the highest membrane damage, which was caused by essential oil (IC50 and IC70), A549 cells had the highest DNA damage at IC50 and IC70 p-cymene concentrations. G6PD, GST, GRx, and GPx enzyme activities of cells, which increased against these compounds, depended on concentrations, incubation times, and antioxidant capacities of the cells. Conclusion: This study suggests that different cancer cells showed different cellular responses against potential antitumor and prooxidative effects of the essential oil and its two main constituents.

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1. Bauer KD, Garbe H, Surburg H. Common Fragrance and Flavor Materials. Weinheim, Germany: Wiley-VCH Verlag; 1997.
2. Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils a review. Food Chem Toxicol 2008; 46: 446-475.
3. Moteki H, Hibasami H, Yamada Y, Katsuzaki H, Imai K, Komiya T. Specific induction of apoptosis by 1,8-cineole in two human leukemia cell lines, but not in human stomach cancer cell line. Oncol Rep 2002; 9: 757-760.
4. Seyoum A, Asres K, El-Fiky FK. Structureradical scavenging activity relationship of flavonoids. Phytochemistry 2006; 67: 2058-2070.
5. Ozkan A, Erdogan A, Sokmen M, Tugrulay S, Unal O. Antitumoral and antioxidant effect of essential oils and in vitro antioxidant properties of essential oils and aqueous extracts from Salvia pisidica. Biologia 2010; 65: 990-996.
6. Erdogan A, Ozkan A. Effects of Thymus revolutus Célak essential oil and its two major constituents on Hep G2 cells membrane. Biologia 2013; 68: 105-111.
7. Afoulous S, Ferhout H, Raoelison EG, Valentin A, Moukarzel B, Couderc F, Bouajila J. Chemical composition and anticancer, antioxidant and antimalarial activities of leaves essential oil of Cedrelopsis grevei. Food Chem Toxıcol 2013; 56: 352-362.
8. Ozkan A, Erdogan A. A comparative study of the antioxidant/ prooxidant effects of carvacrol and thymol at various concentrations on membrane and DNA of parental and drug resistant H1299 cells. Nat Prod Commun 2012; 7: 1557-1560.
9. Sang S, Hou Z, Lambert JD, Yang CS. Redox properties of tea polyphenols and related biological activities. Antioxid Redox Signal 2005; 7: 1704-1714.
10. Nicol CJ, Zielenski J, Tsui LC, Wells PG. An embryoprotective role for glucose-6-phosphate dehydrogenase in developmental oxidative stress and chemical teratogenesis. FASEB J 2000; 14: 111-127.
11. Özkan A, Fışkın K. Epirubicin-HCI cytotoxicity in non-small cell lung cancer (NSCLC) cells. Turk J Hematol Oncol 2003; 13: 125-133.
12. Rihova B, Strohalm J, Kobackova K. Acquired and specific immunological mechanisms co-responsible for efficacy of polymer-bound drugs. J Control Release 2002; 17: 97-114.
13. Morelli MP, Cascone T, Troiani T, De Vita F, Orditura M, Laus G, Eckhardt SG, Pepe S, Tortora G, Ciardiello F. Sequencedependent antiproliferative effects of cytotoxic drugs and epidermal growth factor receptor inhibitors. Ann Oncol 2005; 16 (Suppl. 4): iv61-iv68.
14. Ciardiello F, Tortora G. A novel approach in the treatment of cancer: targeting the epidermal growth factor receptor. Clin Cancer Res 2001; 7: 2958-2970.
15. Mendelsohn J, Baselga J. Status of epidermal growth factor receptor antagonists in the biology and treatment of cancer. J Clin Oncol 2003; 21: 2787-2799.
16. Grunwald V, Hidalgo M. Developing inhibitors of the epidermal growth factor receptor for cancer treatment. J Natl Cancer Inst 2003; 95: 851-867.
17. Jansson O, Motlagh PB, Persson M, Henrikson R, Grankvist K. Increase in doxorubicin cytotoxicity by carvedilol inhibition of P-glycoprotein activity. Biochem Pharmacol 1999; 58: 1801- 1806.
18. Ozkan A. Lymphokine-activated killer cell susceptibility in epirubicin resistant and parental human non-small cell lung cancer (NSCLC). Biologia 2007; 62: 232-237.
19. Gloeckner H, Jonuleit T, Lemke HD. Monitoring of cell viability and cell growth in a hollow-fiber bioreactor by use of the dye Alamar Blue (TM). J Immunol Methods 2001; 252: 131-138.
20. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65: 55-63.
21. Wasowicz W, Neve J, Peretz A. Optimized steps in fluorometric determination of thiobarbituric acid-reactive substances in serum; importance of extraction pH and influence of sample preservation and storage. Clin Chem 1993; 39: 2522-2526.
22. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 1976; 72: 248-254.
23. Invitrogen. Genomic DNA Mini Kit Manual. Carlsbad, CA, USA: Invitrogen; 2012.
24. Garçon G, Dagher Z, Zerimech F, Ledoux F, Courcot D, Aboukais A, Puskaric E, Shirali P. Dunkerque city air pollution particulate matter-induced cytotoxicity, oxidative stress and inflammation in human epithelial lung cells (L132) in culture. Toxicol in Vitro 2006; 20: 519-528.
25. Habig WH, Jakoby WB. Assay for differentiation of glutathione S-transferases. Methods Enzymol 1981; 77: 398-405.
26. Flohe L, Gunzler WA. Glutathione peroxidase. Method Enzymol 1984; 105: 115-121.
27. Carlberg I, Mannervik B. Glutathione reductase. Method Enzymol 1985; 113: 484-499.
28. Buch AD, Archana G, Naresh Kumar G. Enhanced citric acid biosynthesis in Pseudomonas fluorescens ATCC 13525 by overexpression of the Escherichia coli citrate synthase gene. Microbiology 2009; 155: 2620-2629.
29. Kirkman TW. Statistics to Use. Collegeville, MN, USA: Saint Johns University; 1996.
30. Dandlen SA, Lima AS, Mendes MD, Miguel MG, Faleiro ML, Sousa MJ, Pedro LG, Barroso JG, FigueiredoAC. Antimicrobial activity, cytotoxicity and intracellular growth inhibition of Portuguese Thymus essential oils. Rev Bras Farmacogn 2011; 21: 1012-1024.
31. Sertel S, Eichhorn T, Plinkert PK, Efferth T. Cytotoxicity of Thymus vulgaris essential oil towards human oral cavity squamous cell carcinoma. Anticancer Res 2011; 31: 81-87.
32. Özkan A, Erdoğan A. A comparative evaluation of antioxidant and anticancer activity of essential oil from Origanum onites (Lamiaceae) and its two major phenolic components. Turk J Biol 2011; 35: 735-742.
33. Özkan A, Erdoğan A. Membrane and DNA damaging/ protective effects of eugenol, eucalyptol, terpinen-4-ol, and camphor at various concentrations on parental and drugresistant H1299 cells. Turkish J Biol 2013; 37: 405-413.
34. Turna G, Kılıç N, Yıldırım Z, Sarı S. Effects of Thymus sipyleus and taurine on hepatic MDA, GSH, AOPP levels and SOD activity in Ehrlich acide solid tumor model generated mice. Turkiye Klinikleri J Med Sci 2011; 3: 1153-1159 (in Turkish with English abstract).
35. Esmaeili MA, Sonboli A, Mirjalili MH. Oxidative stress protective effect of Dracocephalum multicaule essential oil against human cancer cell line. Nat Prod Res 2014; 28: 848-852.
36. Tang EL, Rajarajeswaran J, Fung SY, Kanthimathi MS. Antioxidant activity of Coriandrum sativum and protection against DNA damage and cancer cell migration. BMC Complement Altern Med 2013; 13: 347.
37. Manjamalai A, Berlin Grace VM. Antioxidant activity of essential oils from Wedelia chinensis (Osbeck) in vitro and in vivo lung cancer bearing C57BL/6 mice. Asian Pac J Cancer Prev 2012; 13: 3065-3071.
38. Bruni R, Medici A, Andreotti E, Fantin C, Muzzoli M, Dehesa M. Chemical composition and biological activities of Ishpingo essential oil, a traditional Ecuadorian spice from Ocotea quixos (Lam.) Kosterm. (Lauraceae) flower calices. Food Chem 2003; 85: 415-421.
39. Sacchetti G, Maietti S, Muzzoli M, Scaglianti M, Manfredini S, Radice M, Bruni R. Comparative evaluation of 11 essential oils of different origin as functional antioxidants, antiradicals and antimicrobials in food. Food Chem 2005; 91: 621-632.
40. Ryu SH, Lee SW, Yang YJ, Song SY, Kim JH, Choi EK, Ahn SD. Intracytoplasmic epidermal growth factor receptor shows poor response to the cetuximab antitumor effect in irradiated nonsmall cell lung cancer cell lines. Lung Cancer 2012; 77: 482- 487.
41. Tian WN, Braunstein LD, Pang J, Stuhlmeier KM, Xi QC, Tian X, Stanton RC. Importance of glucose-6-phosphate dehydrogenase activity for cell growth. J Biol Chem 1998; 273: 10609-10617.
42. De Oliveira JMPF, Costa M, Pedrosa T, Pinto P, Remedios C, Oliveira H, Pimentel F, Almeida L, Santos C. Sulforaphane induces oxidative stress and death by p53-independent mechanism: implication of impaired glutathione recycling. PLoS One 2014; 9: e92980.
43. Tirkey N, Pilkhwal S, Kuhad A, Chopra K. Hesperidin, a citrus bioflavonoid, decreases the oxidative stress produced by carbon tetrachloride in rat liver and kidney. BMC Pharmacol 2005; 5: 2.
44. Koparal AT, Zeytinoglu M. Effects of carvacrol on a human nonsmall cell lung cancer (NSCLC) cell line, A549. Cytotechnology 2003; 43: 149-154.
45. Janzowski C, Glaab V, Samimi E, Schlatter J, Eisenbrand G. 5-Hydroxymethylfurfural: assessment of mutagenicity, DNAdamaging potential and reactivity towards cellular glutathione. Food Chem Toxıcol 2000; 38: 801-809.
46. Munari CC, Oliveira de PF, Souza de Lima IM, de Paula Lima Martins S, de Carvalho da Costa J, Bastos JK, Tavares DC. Evaluation of cytotoxic, genotoxic and antigenotoxic potential of Solanum lycocarpum fruits glicoalkaloid extract in V79 cells. Food Chem Toxicol 2012; 50: 3696-3701.