Dialil Disülfit ve Dialil Trisülfit’in İnsan Prostat Kanser Hücreleri Üzerine Sitotoksik, Genotoksik ve Apoptotik Etkileri

Yıl 2020, , 266 - 277, 20.08.2020

Eray Metin Güler , Hatice Hira Hümeyra Kaleli Abdurrahim Koçyiğit

https://doi.org/10.35440/hutfd.749554

Öz

Amaç: Kanser dünya çapında artan bir sağlık sorunu olup, erkeklerde en sık görülen kanser türü olan prostat kanseri, birçok ülke için ikinci ölüm nedenidir. Kanser tedavisinde konvansiyonel yöntemlerin başarısız olması nedeni ile doğal etken maddelerin kullanımı son yıllarda giderek daha fazla ilgi görmektedir. Bu çalışmanın amacı, Sarımsak (Allium sativum) etken maddelerinden olan, Dialil Disülfit (DADS) ve Dialil Trisülfit’in (DATS) farklı konsantrasyonlarının insan prostat kanser hücreleri üzerine sitotoksik, genotoksik ve apoptotik etkilerini araştırmaktır.
Materyal ve Metot: Bu çalışmada prostat kanser (PC-3) ve sağlıklı prostat epitel hücrelerine (WPMY-1) DADS ve DATS’ın sitotoksik etkisi luminometrik ATP testiyle, genotoksik etkisi alkalen tekli hücre jel elektroforez (Comet Assay) yöntemiyle, apoptotik etkisi akridin turuncusu/etidyum bromür yöntemiyle ölçüldü. Ayrıca, mitokondriyal membran potansiyeli (MMP), hücre içi kalsiyum (Ca2+) ve reaktif oksijen türlerinin (ROS) seviyeleri farklı florometrik yöntemlerle ve glutatyon seviyeleri ise luminometrik yöntem ile tayin edildi.
Bulgular: DADS ve DATS doza bağımlı olarak hem kanser hem de normal hücrelerde glutatyon ve MMP seviyelerini anlamlı şekilde düşürürken, sitotoksisite, DNA hasarı, apoptoz, hücre içi Ca2+ ve ROS düzeylerini anlamlı derecede arttırmıştır. Ayrıca, DATS’ın kanser hücreleri üzerine sitotoksik, genotoksik ve apoptotik etkileri sağlıklı hücrelere ve DADS’a göre daha yüksek bulunmuştur.
Sonuç: Bulgular, hem DADS hem de DATS’ın prostat kanseri hücrelerinde doza bağlı bir şekilde sitotoksik, genotoksik ve apoptotik etkilere sahip olduğunu ve DATS’ın DADS’a göre daha etkili olduğunu göstermiştir. Bu nedenle, DATS’ın prostat kanseri tedavisi için kullanılabilecek seçeneklerden biri olabileceğini önermekteyiz.

Anahtar Kelimeler

Prostat kanseri, dialil disülfit, dialil trisülfit, kanser tedavisi

Destekleyen Kurum

Bezmialem Vakıf Üniversitesi Bilimsel Araştırma Projeleri Birimi

Proje Numarası

6.2016/35 nolu projesi ile desteklenmiştir.

Teşekkür

Bu çalışma 14 Mart 2017’de Bezmialem Vakıf Üniversitesi “Annual Medical Students Research Presentation Day” de kısa sözlü bildiri olarak sunulmuştur.

Cytotoxic, Genotoxic and Apoptotic Effects of Diallyl Disulfide and Diallyl Trisulfide on Human Prostate Cancer Cells

Öz

Abstract
Background: Cancer is an increasing health problem worldwide; however, prostate cancer is the most common cancer in men and the second leading cause of death in many countries. Due to the failure of conventional methods in the treatment of cancer, the use of natural active ingredients in cancer treatment has gained increasing attention in recent years. The aim of this study is to investigate cytotoxic, genotoxic, and apoptotic effects of different concentrations of diallyl disulfide (DADS) and diallyl trisulfide (DATS) on human prostate cancer cells.
Methods: In this study, the cytotoxic effect of DADS and DATS in prostate cancer (PC-3) and healthy prostate epithelial cells (WPMY-1) was measured by luminometric ATP test, the genotoxic effect was measured by alkaline single-cell electrophoresis (Comet Assay) method, and apoptotic effect by acridine orange/ethidium bromide method. In addition, mitochondrial membrane potential (MMP), intracellular calcium (Ca2+), and reactive oxygen species (ROS) levels were measured by different fluorometric methods, and glutathione levels were determined by the luminometric method.
Results: DADS and DATS reduced statistically significantly glutathione and MMP levels while increased cytotoxicity, DNA damage, apoptosis, intracellular Ca2+, and ROS levels in both cancer and healthy cells dose-dependent manner. In addition, the cellular effects of DATS were higher than DADS in cancer cells than healthy cells.
Conclusions: The findings showed that both DADS and DATS have a dose-dependent cytotoxic, genotoxic and apoptotic effect in prostate cancer cells and that DATS is more effective than DADs. Therefore, we suggest that DATS may be one of the options available for the treatment of prostate cancer.

Anahtar Kelimeler

Prostate cancer, diallyl disulfide, diallyl trisulfide, cancer treatment

Proje Numarası

6.2016/35 nolu projesi ile desteklenmiştir.

Kaynakça

• 1. Pienta KJ, Smith DC. Advances in Prostate Cancer Chemotherapy: A New Era Begins 1. CA: a cancer journal for clinicians. 2005;55(5):300-18. 2. Leyh-Bannurah S-R, Gazdovich S, Budäus L, Zaffuto E, Briganti A, Abdollah F, et al. Local therapy improves survival in metastatic prostate cancer. European urology. 2017;72(1):118-24. 3. Mukherjee AK, Basu S, Sarkar N, Ghosh AC. Advances in cancer therapy with plant based natural products. Current medicinal chemistry. 2001;8(12):1467-86. 4. Ross SA, Milner JA. Garlic: the mystical food in health promotion. Handbook of nutraceuticals and functional foods: CRC Press; 2016. p. 84-110. 5. Iciek M, Kwiecień I, Włodek L. Biological properties of garlic and garlic‐derived organosulfur compounds. Environmental and molecular mutagenesis. 2009;50(3):247-65. 6. Cao G, Sofic E, Prior RL. Antioxidant capacity of tea and common vegetables. Journal of agricultural and food chemistry. 1996;44(11):3426-31. 7. Nagini S. Cancer chemoprevention by garlic and its organosulfur compounds-panacea or promise? Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents). 2008;8(3):313-21. 8. Kaschula CH, Hunter R, Parker MI. Garlic‐derived anticancer agents: Structure and biological activity of ajoene. Biofactors. 2010;36(1):78-85. 9. Harris J, Cottrell S, Plummer S, Lloyd D. Antimicrobial properties of Allium sativum (garlic). Applied microbiology and biotechnology. 2001;57(3):282-6. 10. Hosono T, Fukao T, Ogihara J, Ito Y, Shiba H, Seki T, et al. Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of β-tubulin. Journal of Biological Chemistry. 2005;280(50):41487-93. 11. Tan H, Ling H, He J, Yi L, Zhou J, Lin M, et al. Inhibition of ERK and activation of p38 are involved in diallyl disulfide induced apoptosis of leukemia HL-60 cells. Archives of pharmacal research. 2008;31(6):786. 12. Kocyigit A, Guler EM, Karatas E, Caglar H, Bulut H. Dose-dependent proliferative and cytotoxic effects of melatonin on human epidermoid carcinoma and normal skin fibroblast cells. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2018;829:50-60. 13. Kocyigit A, Guler EM, Karatas E, Caglar H, Bulut H. Dose-dependent proliferative and cytotoxic effects of melatonin on human epidermoid carcinoma and normal skin fibroblast cells. Mutation research. 2018;829-830:50-60. 14. Untario N, Dewi TC, Widodo MA, Rahaju P. Effect of Tetrodotoxin from Crude Puffer Fish (Tetraodon fluviatilis) Liver Extract on Intracellular Calcium Level and Apoptosis of HeLa Cell Culture. Journal of Tropical Life Science. 2017;7(1):23-9. 15. Rottenberg H, Wu S. Quantitative assay by flow cytometry of the mitochondrial membrane potential in intact cells. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research. 1998;1404(3):393-404. 16. Günes-Bayir A, Kiziltan HS, Kocyigit A, Güler EM, Karataş E, Toprak A. Effects of natural phenolic compound carvacrol on the human gastric adenocarcinoma (AGS) cells in vitro. Anti-cancer drugs. 2017;28(5):522-30. 17. Singh NP, Danner DB, Tice RR, Brant L, Schneider EL. DNA damage and rpair with age in individual human lymphocytes. Mutation research. 1990;237(3-4):123-30. 18. Demirbag R, Yilmaz R, Gur M, Kocyigit A, Celik H, Guzel S, et al. Lymphocyte DNA damage in patients with acute coronary syndrome and its relationship with severity of acute coronary syndrome. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis. 2005;578(1-2):298-307. 19. Kasibhatla S, Amarante-Mendes GP, Finucane D, Brunner T, Bossy-Wetzel E, Green DR. Acridine Orange/Ethidium Bromide (AO/EB) Staining to Detect Apoptosis. CSH protocols. 2006;2006(3). 20. Komura K, Sweeney CJ, Inamoto T, Ibuki N, Azuma H, Kantoff PW. Current treatment strategies for advanced prostate cancer. International Journal of Urology. 2018;25(3):220-31. 21. Song Y-h, Sun H, Zhang A-h, Yan G-l, Han Y, Wang X-j. Plant-derived natural products as leads to anti-cancer drugs. J Med Plant Herb Ther Res. 2014;2:6-15. 22. Schumacker PT. Reactive oxygen species in cancer cells: live by the sword, die by the sword. Cancer cell. 2006;10(3):175-6. 23. Ling H, Zhang L-Y, Su Q, Song Y, Luo Z-Y, Zhou XT, et al. Erk is involved in the differentiation induced by diallyl disulfide in the human gastric cancer cell line MGC803. Cellular & molecular biology letters. 2006;11(3):408. 24. Knowles L, Milner J. Diallyl disulfide induces ERK phosphorylation and alters gene expression profiles in human colon tumor cells. The Journal of nutrition. 2003;133(9):2901-6. 25. Lai KC, Hsu SC, Kuo CL, Yang JS, Ma CY, Lu HF, et al. Diallyl sulfide, diallyl disulfide, and diallyl trisulfide inhibit migration and invasion in human colon cancer colo 205 cells through the inhibition of matrix metalloproteinase‐2,‐7, and‐9 expressions. Environmental toxicology. 2013;28(9):479-88. 26. Druesne-Pecollo N, Pagniez A, Thomas M, Cherbuy C, Duée P-H, Martel P, et al. Diallyl disulfide increases CDKN1A promoter-associated histone acetylation in human colon tumor cell lines. Journal of agricultural and food chemistry. 2006;54(20):7503-7. 27. Xiao D, Pinto JT, Gundersen GG, Weinstein IB. Effects of a series of organosulfur compounds on mitotic arrest and induction of apoptosis in colon cancer cells. Molecular cancer therapeutics. 2005;4(9):1388-98. 28. Yin M-c, Hwang S-w, Chan K-c. Nonenzymatic antioxidant activity of four organosulfur compounds derived from garlic. Journal of Agricultural and Food Chemistry. 2002;50(21):6143-7. 29. Wu X-J, Kassie F, Mersch-Sundermann V. The role of reactive oxygen species (ROS) production on diallyl disulfide (DADS) induced apoptosis and cell cycle arrest in human A549 lung carcinoma cells. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis. 2005;579(1-2):115-24. 30. Yordi EG, Pérez EM, Matos MJ, Villares EU. Antioxidant and pro-oxidant effects of polyphenolic compounds and structure-activity relationship evidence. Nutrition, well-being and health. 2012;10:29471. 31. Das A, Banik NL, Ray SK. Garlic compounds generate reactive oxygen species leading to activation of stress kinases and cysteine proteases for apoptosis in human glioblastoma T98G and U87MG cells. Cancer. 2007;110(5):1083-95. 32. Liao Y, Bai H, Li Z, Zou J, Chen J, Zheng F, et al. Longikaurin A, a natural ent-kaurane, induces G2/M phase arrest via downregulation of Skp2 and apoptosis induction through ROS/JNK/c-Jun pathway in hepatocellular carcinoma cells. Cell death & disease. 2014;5(3):e1137-e. 33. Berridge MJ. Calcium microdomains: organization and function. Cell Calcium. 2006;40(5-6):405-12. 34. Hajnóczky G, Csordás G. Calcium signalling: fishing out molecules of mitochondrial calcium transport. Curr Biol. 2010;20(20):R888-91. 35. Ly JD, Grubb DR, Lawen A. The mitochondrial membrane potential (Δψ m) in apoptosis; an update. Apoptosis. 2003;8(2):115-28. 36. Kim Y-A, Xiao D, Xiao H, Powolny AA, Lew KL, Reilly ML, et al. Mitochondria-mediated apoptosis by diallyl trisulfide in human prostate cancer cells is associated with generation of reactive oxygen species and regulated by Bax/Bak. Molecular cancer therapeutics. 2007;6(5):1599-609. 37. Truong D, Hindmarsh W, O’Brien P. The molecular mechanisms of diallyl disulfide and diallyl sulfide induced hepatocyte cytotoxicity. Chemico-biological interactions. 2009;180(1):79-88. 38. Choi YH, Park HS. Apoptosis induction of U937 human leukemia cells by diallyl trisulfide induces through generation of reactive oxygen species. Journal of biomedical science. 2012;19(1):50. 39. Lushchak VI. Glutathione homeostasis and functions: potential targets for medical interventions. Journal of amino acids. 2012;2012. 40. Sielicka-Dudzin A, Borkowska A, Herman-Antosiewicz A, Wozniak M, Jozwik A, Fedeli D, et al. Impact of JNK1, JNK2, and ligase Itch on reactive oxygen species formation and survival of prostate cancer cells treated with diallyl trisulfide. European journal of nutrition. 2012;51(5):573-81.