Effects of essential oils obtained from wild and cultured forms of thyme (Origanum acutidens) on lung cancer cells membrane

Abstract

Aim: Nowadays, studies on the use of medicinal plants and their essential oils in cancer treatment are increasing rapidly and these studies are very important both scientifically and economically. However, the collection of plants from nature causes extinction. Therefore, it is of great importance to cultivate plants, especially endemic species, to reproduce and consume them. In our study, cytotoxic and membrane damaging effects of essential oils obtained from wild and cultured forms of Origanum acutidens (Hand.-Mazz.) Ietswaart (Lamiaceae) (O. acutidens), which is endemic to Turkey, on non-small-cell lung cancer (NSCLC) cells H1299 and A549 were compared. Material and Method: The level of malondialdehyde, an oxidative stress biomarker, was determined in cell lysates. Assessment of cell viability was made by CellTiter-Blue® Cell Viability Assay and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay after 10- 250 μg/mL concentrations of wild and cultured forms of O. acutidens essential oil treated to H1299 and A549 cells for 24, 48 and 72 h. Malondialdehyde levels were assayed for determining the membrane damaging effects. Results: Cell viability of H1299 and A549 cells incubated with essential oils obtained from wild and cultured forms of O. acutidens was found to decrease depending on concentration and time. 179 µg/mL, 157 µg/mL, and 132 µg/mL were calculated as IC50 values of wild form of O. acutidens essential oil on H1299 cells for 24, 48, and 72 h, respectively by MTT assay. 150 µg/mL, 131 µg/mL, and 110 µg/mL were calculated as IC50 values of wild form of O. acutidens essential oil on H1299 cells for 24, 48, and 72 h, respectively by resazurin-based assay. 118 µg/mL, 99 µg/mL, and 69 µg/mL were calculated as IC50 values of wild form of O. acutidens essential oil on A549 cells for 24, 48, and 72 h, respectively by MTT assay. 98 µg/mL, 83 µg/mL, and 57 µg/mL were calculated as IC50 values of wild form of O. acutidens essential oil on A549 cells for 24, 48, and 72 h, respectively by resazurin-based assay. Essential oils obtained from wild and cultured forms of O. acutidens increased malondialdehyde level on both H1299 and A549 cells. The greatest membrane damage was observed in A549 cells treated with wild form of O. acutidens essential oil. Conclusion: Essential oils obtained from wild and cultured forms of O. acutidens had cytotoxic effect on lung cancer cells and it has been demonstrated that they showed this effect by causing membrane damage in cells.

Keywords

• O. acutidens
• essential oil
• wild and cultured form
• membrane damaging
• anticancer

Thanks

The authors wish to thanks to Alanya Alaaddin Keykubat University for providing the necessary facilities to conduct this study.

References

1. Yaşar S. Determination of soil properties, fixed and essential oil contents of some perennial medicinal plants that grow naturally in Çukurova University campus. Master Thesis, Çukurova University 2005; 43 pp.
2. World Cancer Research Fund International. Working to prevent cancer worldwide. Available online: www.wcrf.org, 2012
3. Iscan M. The role of metabolic polymorphism in drug resistance in lung cancer. Ankara University Scientific Research Projects, 2007, Ankara.
4. Cragg GM, Newman DJ. Antineoplastic agents from natural sources: achievements and future directions. Expert Opin Investig Drugs 2000; 9: 1-15.
5. Usta J, Kreydiyyeh S, Knio K, Barnabe P, Bou-Moughlabay Y, Dagher S. Linalool decreases Hep G2 viability by inhibiting mitochondrial complexes I and II, increasing reactive oxygen species and decreasing ATP and GSH levels. Chem-Biol Interact 2009; 180: 39-46.
6. 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: 131-8.
7. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65: 55-63.
8. Wasowicz W, Neve J, Peretz A. Optimized steps in fluorometric determination of thiobarbitiric acidreactive substances in serum; importance of extraction pH and influence of sample preservation and storage. Clin Chem 1993; 39: 2522-6.

9. 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: 248-54.
10. Kirkman TW. Statistics to use [Online]. 1996 Available:http://www.physics.csbsju.edu/stats/1996 [17 August 2008].
11. Griffiths DE. Psi-screen, an in vitro toxicity test system: applications in the bioassay of perfumes and fragrance chemicals. Altern Lab Anim 2005; 33: 471-86.
12. Lambert RJV, Skandamis PN, Coote PJ, Nychas GJ. A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol. J Appl Microbiol 2001; 91: 453-62.
13. Ozkan A, Erdogan 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: 735-42.
14. Erdogan A, Ozkan A. A comparative study of cytotoxic, membrane and DNA damaging effects of Origanum majorana’s essential oil and its oxygenated monoterpene component linalool on parental and epirubicin-resistant H1299 cells. Biologia 2013; 68: 754-61.
15. Sobral MV, Xavier AL, Lima TC, De Sousa DP. Antitumor activity of monoterpenes found in essential oils. Hindawi Publishing Corporation, Scientific World Journal 2014; Article ID 953451: 35 pages.
16. Bayala B. Anticancer activity of essential oils and their chemical components -a review. Am J Cancer Res 2014; 4: 591-607.
17. Essien EE, Newby JM, Walker TM, Ogunwande IA, Setzer WN, Ekundayo O. Essential oil constituents, anticancer and antimicrobial activity of Ficus mucoso and Casuarina equisetifolia leaves. J Essent Oils 2016; 41: 1-6.
18. Tuncer E, Unver-Saraydın S, Tepe B, et al. Antitumor effects of Origanum acutidens extracts on human breast cancer. J Buon 2013; 18: 77-85.
19. Turan M, Sökmen A, Karadayı K, Polat ZA, Şen M. Anti-neoplastic effects of some plant extracts specific to Sivas region. Cumhur Medical J 2010; 32: 9-18.
20. Kursat M, Emre İ, Yılmaz Ö, Erecevit P. Antioxidant and antimicrobial activity in the seeds of Origanum vulgare L. subsp. gracile (C. Koch) Ietswaart and Origanum acutidens (Hand.-Mazz.) Ietswaart from Turkey. Grasas Aceites 2011; 62: 410.
21. Osge B, Turker A, Ipek A, Gurbuz B. Chemical compositions and antibacterial activities of the essential oils from aerial parts and corollas of Origanum acutidens (Hand.-Mazz.) Ietswaart, an endemic species to Turkey. Molecules 2009; 14: 1702-12.
22. Ozkan A, Erdogan 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: 735-42.
23. Ozkan A, Erdogan 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: 405-13.
24. 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: 990-96.
25. Erdogan A, Ozkan A. Effects of Thymus revolutus Célak essential oil and its two major components on Hep G2 cells membrane. Biologia 2013; 68: 105-11.