Feyzan Özdal KURT, H. Seda VATANSEVER, Kadriye SORKUN, S. Ismet Delıloğlu GURHAN, ELGİN TÜRKÖZ ULUER, Ömür GENÇAY, Bekir SALİH

Inhibitory effects of Propolis on human osteogenic sarcoma cell proliferation mediated by caspase patway

Doğal bir ürün olarak propolis belli ağaç ve bitkilerin kabuki ve tomurcuklarından arılar aracılığı ile elde edilen bir reçine materyalidir. Propolis antimutajenik, antioksidan, antibakteriyel, antiviral etkileride içeren biyolojik aktiviteleri olan çeşitli kimyasal bileşikleri içerir. Bu nedenle bu çalışmada insan osteojenik sarcoma hücre dizini SAOS-2 kültüründe kaspaz yolağı kullanılarak propolis ekstraktlarının (PE) antiapoptotik etkisi araştırılması amaçlandı. Ankara’da Hacettepe Üniversitesi Beytepe Kampüs alanında ekolojik çevrede üretilen ekstraktlar kullanıldı. 0.5, 0.25, 0.125 ve 0.063 mg/ml konsantrasyonlarda 7 farklı PE SAOS-2 hücre dizinine eklenerek 2 gün boyunca inkübe edildi. Hücre çoğalması ve toksisitesi için MTT testi ile apoptotic hücre ölümünün belirlenmesi için TUNEL metodu ve kaspaz dağılımı için kaspaz 6, 8 ve 9 immunohistokimyası analizi kullanıldı. Hücreler MTT analizi sonrasında en büyük etki PE 7 için 0.125 mg/ml dilüsyonunda görüldü. TUNEL pozitif hücre sayısı en fazla olan PE 4 ve 5 için 0.125 mg/ml dilüsyonu oldu. Kaspaz 6 immünoreaktivitesi kaspaz 8 ve 9’dan daha güçlü idi. Daha da önemlisi kaspaz 6 boyaması PE 7 için 0.125 mg/ml dilüsyonunda çok daha iyi idi. Sonuç olarak, PE ile indüklenen apoptosis mekanizmasının antikansorejenik etkisinden dolayı kaspaz yolağı ile oluşabilir. PE kullanımı kanser tedavi protokolünde yararlı olabilir.

Anahtar Kelimeler:

sarkom, programlanmış hücre ölümü, hücre hatları, sitotoksisite, enzimler, insan, neoplazma, propolis

Kaspaz yolağı ile yönlendirilen insan osteojenik sarcoma hücre çoğalmasına Propolisin baskılayıcı etkisi

A natural product Propolis, is a resinous material gathered by honeybees from the buds and bark of certain trees and plants. Propolis contains various chemical components of biological activities, including antimutagenic, antioxidant, antibacterial, antiviral and anticarsinogenic. Therefore, the aim of this study is to investigate the antiapoptotic effect of propolis extracts (PE) using caspase pathway in the human osteogenic sarcoma cell line SAOS-2 in culture. The extracts which produced in ecologic environment were taken from the Hacettepe University, Beytepe Campus area-Ankara were used. Seven different PE at 0.5, 0.25, 0.125 and 0.063 mg/ml were added to SAOS-2 cell line for two days incubation. For cell proliferation and cytotoxicty analyses MTT, for apoptotic cell death determination TUNEL method, for distribution of caspase 6, caspase 8 and caspase 9 indirect immunocytochemistry analyses were used. After MTT analyses, the most effect was observed PE 7 at the 0.125 mg/ml dilution. The number of TUNEL positive cells was more detectable at PE 4 and 5 at the 0.063 mg/ml, and PE 7 at the 0.125 mg/ml dilutions. The immunoreactivity of caspase 6 was stronger than caspase 8 and 9. Moreover, density of caspase 6 staining was much better especially in PE 7 at the 0.125 mg/ml dilution. In conclusion, the mechanisms of apoptosis induction by PE may appear via caspase pathway because of its anticanserogenic effect. PE may be usefull in the cancer treatment protocol.

Keywords:

sarcoma, apoptosis, cell lines, cytotoxicity, enzymes, man, neoplasms, propolis,

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1.Sorkun K, Suer B, Salih B: Determination of chemical composition of Turkish propolis. Z Naturforsch, 56, 666-668, 2001.
2.Kartal M, Yıldız S, Kaya S, Kurucu S, Topçu G: Antimicrobial activity of propolis samples from two different regions of Anatolia. J Ethnopharmacol, 86, 69-73, 2003.
3.Isla MI, Moreno Nieva MI, Sampietro AR, Vattuone MA: Antioxidant activity of Argentine propolis extracts. J Ethnopharmacol, 76, 165-170, 2001
4.Padmavathi R, Senthilnathan P, Sakthisekaran D: Therapeutic effect of propolis and paclitaxel on hepatic phase I and II enzymes and marker enzymes in dimethylbenz(a)anthraceneinduced breast cancer in female rats. Comp Biochem Physiol C Toxicol Pharmacol, 143 (3): 349-354, 2006.
5.Bankova VS, Popov SS, Marekov NL: Isopentenyl cinnamates from poplar buds and propolis. Phytochem, 28, 871-873, 1989.
6.Schwartzman RA, Cidlowski JA: Apoptosis: The biochemistry and molecular biology of programmed cell death. Endocrinol Rev, 14, 133-151, 1993.
7.Green DR, Reed JC: Mitochondria and apoptosis. Sci, 281, 1308-1312, 1998.
8.Stellar H: Mechanisms and genes of cellular suicide. Sci, 267, 1445-1449, 1995.
9.Nagata S, Golstein P: The Fas death factor. Sci, 267, 14491456, 1995.
10.Okura T, Gang L, Kamitami T, Wada T, Okura I, Wei CF, Chang HM, Yeh ET: Protection against Fas/Apo-1 and tumor necrosis factor-mediated cell death by a novel protein, sentrin. J Immunol, 157, 4277-4281, 1996.
11. Sattler M, Liang H, Nettesheim D, Meadows RP, Harlan JE, Eberstadt M, Yoon HS, Shuker SB, Chang BS, Minn AJ, Thompson CB, Fesik SW: Structure of Bcl-xL-Bak peptide complex: Recognition between regulators of apoptosis. Sci, 275, 983-986, 1997.
12. Thornberry NA, Lazebnik Y: Caspases: Enemies within. Sci, 281, 1312-1316, 1998.
13. Wyllie AH, Bellamy CO, Bubb VJ, Clarke AR, Corbet S, Curtis L, Harrison DJ, Hooper ML, Toft N, Webb S, Bird CC: Apoptosis and carcinogenesis. Br J Cancer, 80 Supp l, 34-37, 1999.
14. Reed JC: Mechanisms of apoptosis avoidance in cancer. Curr Opin Oncol, 11, 68-75, 1999.
15. Igney FH, Krammer PH: Death and anti-death: Tumour resistance to apoptosis. Nat Rev Cancer, 2, 277-288, 2002.
16. Chen JH, Shao Y, Huang MT, Chin CK, Ho CT: Inhibitory effect of caffeic acid phenethyl ester on human leukemia HL60 cells. Cancer Lett, 108, 211-214, 1996.
17. Aso K, Kanno S, Tadano T, Satoh S, Ishikawa M: Inhibitory effect of propolis on the growth of human leukemia U937. Biol Pharm Bull, 27, 727-730, 2004.
18. Chen CN, Wu CL, Lin JK: Propolin C from propolis induces apoptosis through activating caspases, Bid and cytochrome c release in human melanoma cells. Biochem Pharmacol, 67, 53-66, 2004.
19. Chen CN, Wu CL, Lin JK: Apoptosis of human melanoma cells induced by the novel compounds propolin A and propolin B from Taiwenese propolis. Cancer Lett, 245, 218231, 2007.
20. Deliloglu-Gürhan SI, Vatansever HS, Özdal-Kurt F, Tuğlu I: Characterization analysis of osteoblast derived from bone marrow in modified culture system. Acta Histochem, 108, 4957, 2006.
21. Vatansever HS, Sorkun K, Deliloglu-Gürhan SI, Özdal-Kurt F, Turkoz E, Gencay O, Salih B: Propolis from Turkey induces apoptosis through activating caspases in human breast carcinoma cell lines. Acta Histochem, 3, (Epub ahead of print), 2009.
22. Ergün G, Aktaş S: ANOVA modellerinde kareler toplamı yöntemlerinin karşılaştırılması. Kafkas Univ Vet Fak Derg, 15 (3): 481-484, 2009.
23. Marucci MC: Propolis: Chemical composition, biological properties and therapeutic activity. Apidologie, 26, 83-99, 1995.
24. Khalil ML: Biological activity of bee propolis in health and disease. Asian Pac J Cancer Prev, 7 (1): 22-31, 2006.
25. Matsukawa Y, Marui N, Sakai T, Satomi Y, Yoshida M, Matsumoto K: Genistein arrests cell cycle progression at G2M. Cancer Res, 53, 1328-1331, 1993.
26. Reed JC: Mechanisms of apoptosis. Am J Pathol, 157, 1415-1430, 2000.
27. Huang WJ, Huang CH, Wu CL, Lin JK, Chen YW, Lin CL, Chuang SE, Huang CY, Chen CN: Propolin G, a prenylflavanone, isolated from Taiwanese propolis, induces caspase-dependent apoptosis in brain cancer cells. J Agric Food Chem, 55 (18): 7366-7376, 2007.
28. Li H, Kapur A, Yang JX, Srivastava S, McLeod DG, Paredes-Guzman JF, Daugsch A, Park YK, Rhim JS: Anti- proliferation of human prostate cancer cells by ethanolic extracts of Brazilian propolis and its botanical origin. Int J Oncol, 31 (3): 601-606, 2007.
29. Weng MS, Liao CH, Chen CN, Wu CL, Lin JK: Propolin H from Taiwanese propolis induces G1 arrest in human lung carcinoma cells. J Agric Food Chem, 55 (13): 5289-5298, 2007.
30. Özcan A, Atakişi E: Apoptosisin biyokimyasal önemi. Kafkas Univ Vet Fak Derg, 9 (1): 95-100, 2003.
31. Balcan E, Arslan Ö, Gümüş A, Şahin E: Tunikamisinin embriyonik ve yenidoğan fare dalak dokularına etkisi. Kafkas Univ Vet Fak Derg, 15 (5): 651-660, 2009.