BibTex RIS Kaynak Göster

Insan lenfosit hücre kültüründe melatonin ve b-karotenin mitomisin C ile indüklenmiş kardeş kromatid deşişimi sıklışı üzerine in vitro etkileri

Yıl 2008, Cilt: 15 Sayı: 3, 23 - 29, 26.05.2009

Öz

Süleyman Demirel Üniversitesi
TIP FAKÜLTESİ DERGİSİ: 2008 Eylül; 15 (3)


Insan lenfosit hücre kültüründe melatonin ve b-karotenin mitomisin C ile indüklenmiş kardeş kromatid deşişimi sıklışı üzerine in vitro etkileri

Yasemin Soysal*, Feride Iffet Şahin**, Sevda Menevşe***

Özet
Bu çalışmanın amacı, antioksidan etkileri bildirilmiş maddeler melatonin, -karoten ve genotoksik bir madde olan Mitomisin C.nin (MMC) tek tek veya birlikte in vitro ortamda insan periferik kan lenfosit kültürlerinde kardeş kromatid değişimi (KKD) üzerindeki etkilerini araştırmaktır. Sağlıklı 11 bireyin periferik kan lenfosit kültürlerinden Fluorescence Plus Giemsa (FPG) tekniği modifiye edilerek kardeş kromatid değişimi değerleri tespit edilmiştir. Melatoninin ve β-karotenin (beta-karoten) tek başlarına eklendikleri kültürlerle ve madde içermeyen kontrolle karşılaştırıldığında benzer KKD değerlerine sahip oldukları, melatoninin ve –karotenin birlikte kültürlere eklenmeleri sonucu, madde içermeyen kontrol ve tek başlarına kullanıldıkları kültürlerin değerleri ile karşılaştırıldığında KKD değerlerinde farklılık oluşturmadıkları gözlenmiştir. MMC, beklenen KKD deşerini arttırma etkisini göstermiştir. Melatonin ve β-karoten tek başlarına ve birlikte kullanıldıkları durumda MMC.nin KKD.ni indükleyen etkisini azaltmıştır. MMC.nin tek başına kullanımındaki sonuçla karşılaştırıldığında melatonin, β-karoten ve MMC.nin birlikte kullanıldığı kültürde KKD oranı anlamlı düzeyde azalmıştır. MMC.nin KKD oluşumunu indükleyen etkisi karşısında β-karoten, KKD değerlerini melatonine kıyasla daha fazla azaltmış ve melatonin ile karşılaştırıldığında daha güçlü antimutajenik etki gösterdiği saptanmıştır.

Anahtar Sözcükler: kardeş kromatid değişimi, melatonin, beta-karoten, mitomisin C, genotoksik etki

Abstract

The in vitro effect of melatonin and -carotene on sister chromatid exchange frequency in mitomycin C-induced human blood lymphocytes The aim of this study was to investigate the in vitro effects of melatonin, β-carotene (known as antioxidants) and Mitomycin C (MMC) (known as a genotoxic agent) separately and together on sister chromatid exchance
(SCE) in human blood lymphocyte cultures. A total of 11 healthy human donors have been included in this research and their peripheral blood culture cells were processed by modified fluorescence plus Giemsamethod to investigate sister chromatid Exchange frequency. The results indicate that melatonin and β-carotene (beta-carotene) alone and compared to untreated controls have similar incidance of SCE. Melatonin and beta-carotene together in the cell culture did not exhibit significant change in the frequency of SCE compared to the cultures they were alone and untreated controls. The lymphocytes, which were treated with mitomycin C (MMC), showed the expected increased incidence of SCE. Melatonin and
Β-carotene used together and alone decreased the MMC induced frequency of SCE. Melatonin, β-carotene and MMC together have significant decrease of SCE frequency when compared to MMC alone. We observed that, beta-carotene decreases the frequency of SCE more than melatonin. So, we concluded that beta-carotene showed stronger antimutagenic effect than melatonin according to the frequency of sister chromatid exchange.

Key words: sister chromatid exchange, melatonin, beta-carotene, mitomycin C, genotoxic effect

Kaynakça

  • Stich HF, Dunn BP. Relationship between cellular levels of beta-carotene and sensitivity to genotoxic agents. Int J Cancer 1986; 38: 713-717
  • Carrano AV, Natarajan AT. Considerations for population monitoring using cytogenetic techniques. Mutat. Res 1988; 204: 379-406
  • Perry PE, Thomson EJ. The methodology of sister chromatid exchanges. In: Kilbey BJ, Legator M, Nichols W, Ramel C. Handbook of Mutagenicity Test Procedures, Elsevier Science Publishers, 1984; 495- 529
  • Emre S. Antikanser ilaçlarýn ve karsinojen maddelerin insan kromozomlarý üzerine etkilerinin in vitro sistemde kardeþ kromatid deðiþimi (sister chromatid exchange, SCE ) analiz yöntemi ile belirlenmesi. Doktora, Hacettepe Üniversitesi Saðlýk Bilimleri Enstitüsü, Týbbi Biyoloji Anabilim Dalý, Ankara,1989.
  • Van Rensburg CEJ, Theron A, Richards GA, Van der Merwe CA, Anderson R. Investigation of the relationship between plasma levels of ascorbate, vitamin E and -Carotene and the frequency of sister chromatid exchanges and release of reactive oxidants by blood leucocytes from cigarette smookers. Mutat Res 1989; 215: 167-172
  • Manoharan K, Banerjee MR. -Carotene reduces sister chromatid exchanges induced by chemical carcinogens in mouse mammary cells in organ culture. Cell Biology International Reports 1985; 9(9): 783-789
  • Bianchi L, Tateo F, Pizzala R, Stivala LA, Verri GM, Melli R, Santamaria L. Carotenoids reduce the chromosomal damege induced by bleomycin in human cultured lymphocytes. Anticancer Research 1993; 13: 1007-1010
  • Konopack M, Widel M, Rzeszowska-Wolny J. Modifying effect of vitamins C, E and beta-carotene against gamma-ray -induced DNA damage in mouse cells. Mutat Res 1998; 417: 85-94
  • Salvadori DMF, Ribeiro LR, Oliveira MDM, Pereira CAB, Beçak W. The protective effect of beta-carotene on genotoxicity induced by cyclophophamide. Mutat Res 1992; 265: 237-244
  • Bal F, Þahin FÝ, Yirmibeþ M, Balcý A, Menevþe S. The in vitro effect of beta-carotene and mitomycin C on SCE frequency in Down's Syndrome lymphocyte cultures. Tohoku J Exp Med 1998; 184(4): 295-300
  • Yamagata Z, Lijima S, Takashita T, Ariizumi C, Hugurashi M. Mitomycin C induced sister chromatid exchanges and cell cycle kinetics in lymphocytes from patients with Klinefelter Syndrome. Mutat Res 1989; 212: 263-268
  • Holman A, Karlsson A, Bratt I, Högstedt B. Micronuclei and mitotic index in B-, T4- and T8- cells treated with mitomycin C and -Irradiation. Mutat Res 1994; 309: 93-99
  • Vijayalaxmi, Reiter RJ, Leal BZ, Meltz ML. Effect of melatonin on mitotic and proliferation indices, and sister chromatid exchange in human blood lymphocytes. Mutat Res 1996; 351: 187-192
  • Raj AS, Katz M. Beta-carotene as an inhibitor of benzo(a)pyrene and mitomycin C induced chromosomal breaks in the bone marrow of mice. Can J Genet Cytol 1985; 27(5): 598-602
  • Bianchi NO, Bianchi MS, Larramendy M. Kinetics of human lymphocyte division and chromosomal radiosensitivity, Mutat Res 1979; 63: 317-324
  • Chapner BA, Myers CE. Antitumor Antibiotics, Cancer. In: De Vita, VT, Hellman S, Rosenberg SA. Principles and Practice of Oncology, J.B. Lippincott Company, Philadelphia, 1993; Vol.2, 381.
  • Richards GA, Theron AJ, Van Resburg EC, Van Rensburg JA, Van der Merwe CA, Kuyl MJ, Anderson R. Investigation of the effects of oral administration of vitamin E and beta-carotene on the chemiluminescence responses and the frequency of sister chromatid exchanges in circulating leucocytes from cigarette smokers. Am Rev Respir Dis 1990; 142: 648-654
  • Xue K-X, Wu J-Z, Ma G-J, Yuan S, Qiu H-L. Comparative studies on genotoxicity and antigenotoxicity of natural and synthetic beta-carotene stereoisomers. Mutat Res 1998; 418: 73-78.
  • Schwartz JK. In vitro biological methods for determination of carotenoid activity. Methods Enzymol 1993; 214: 226-256
  • Ziegler RG. Vegetables, fruits and carotenoids and the risk of cancer. Am J Clin Nutrition 1991; 53: 251-259
  • Brzezinski A. Melatonin in humans. The New England Journal of Medicine 1997; 336(3): 186-195
  • Pelaez AM, Reiter RJ. Distribution of melatonin in mammalian tissues: the relative importance of nuclear versus cytosolic localization. J Pineal Res 1993; 15: 59-69
  • Ekmekçi A, Þaylý A, Dönmez H, Bal F. In vitro effects of prostaglandin E1 and indomethacin on mitomycin C induced human lymphocytes, Mutat Res 1995; 328: 49-53.
  • Perry PE, Wolff S. New Giemsa method for the differential staining of sister chromatids. Nature 1974; 251-56.
  • Ekmekçi A, Þaylý A. Cytogenetic study of tuberculosis patients before and after tuberculastic drug treatment. Mutat Res 1995; 34: 175-183.
  • Verma RS, Babu A. Human Chromosomes: Priciples and Techniques, (Second Ed) United States of America, McGraw-Hill, Inc., 1995; 143-151.
  • Howell RT, Taylor AHK. Chromosome instability syndromes. In: Rooney DE, Czepulkowski BH. Human Cytogenetics: A Practical Approach, New York, Oxford University Pres, 1992; Vol 2: 209-233
  • WHO. Guide lines fort he study of genetic effects in human populations. Environmental Health Criteria , Geneva, World Health Organisation, 1985; 46:45-55
  • Carrono AV, Natarajan AT. Considerations for population monitoring using cytogenetic techniques. Mutat Res 1988; 204: 379-406
  • De Flora S, Ramel C. Mechanisms of inhibitors of mutagenesis and carcinogenesis, classification and overview. Mutat Res 1988; 202: 285-306
  • Ustundag A, Duydu Y. The influence of melatonin and N-acetylcystein in delta-aminolevulinic acid and lead induced genotoxicity in lymphocytes in vitro. Biol Trace Elem Res 2007; 117(1-3): 53-64
  • Antolin I, Rodriguez C, Sainz RM, Mayo JC, Uria H, Kotler ML, Rodriguez-Colunga MJ, Tolivia D, Menendez-Pelaez A. Neurohormone melatonin prevents cell damage: effect on gene expression for antioxidant enzymes. FASEB J 1996; 10(8): 882-90
  • Rajas E, Montero R, Herrera LA, Sardo M, Gonsebatt ME, Radrigues R, Ostrosky-Wegman P. Are mitotic index and lymphocyte proliferation kinetics reproducible endpoints in genetic toxicology testing? Mutat Res 1992; 282: 283-286.
  • Rachel AJ, Sharma T, Menan VV. Harlequin banding and localisation of sister chromatid exchanges. Mutat Res 1991; 264: 71-80
  • Kopjar N, Miocic S, Ramic S, Milic M, Viculin T. Assessment of the radioprotective effects of amifostine and melatonin on human lymphocytes irradiated with gamma-rays in vitro. Arh Hig Rada Toxikol 2006; 57(2): 155-63
  • Van der Rensburg CEJ, Theron A, Richards GA, Van der Merwe CA, Anderson R. Investigation of the relationship between plasma levels of ascorbate, vitamin E and b-carotene and the frequency of sister chromatide and rRelease of reactive oxidants by blood leucocytes from cigarette smookers. Mutat Res 1989; 215: 167- 172
  • Sarkar A, Baþak R, Bishayee A, J, Chatterjee M. Beta- carotene inhibits rat liver chromosomal aberrations and DNA chain break after a single Injection of diethylnitrosamine. Br J Cancer 1997; 76(7): 855-61
  • Alija AJ, Bresgen N, Sommerburg O, Langhans CD, Siems W, Eckl PM. Cyto-and genotoxic potential of beta-carotene and cleavage products under oxidative stres. Biofactors 2005;24(1-4): 159-63
  • Buyukokuroglu ME, Cemek M, Yurumez Y, Yavuz Y, Aslan A. Antioxidative role of melatonin in organophosphate toxicity in rats. Cell Biol Toxicol 2007; Sep.2, online.
  • Sadir S, Deveci S, Korkmaz A, Oter S. Alpha- tocopherol, beta-carotene and melatonin administration protects cyclophosphamide-induced oxidative damage to bladder tissue in rats. Cell Biochem Funct 2007; 25(5): 521-6
  • Yýldýrým I, Korkmaz A, Oter S, Ozcan A, Oztas E. Contribution of antioxidants to preventive effect of mesna in cyclophosphamide-induced hemorrhagic cystitis in rats. Cancer Chemother Pharmacol 2004; 54(5): 469-73
  • Oyanagui Y. Natural antioxidants enhance and prolong the oxyradical/no-related suppression by dexamethasene of ischemic and histamine paw edame in mice. Inflammation 1997; 21(6): 643- 654
Toplam 42 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Bölüm Araştırma Makaleleri
Yazarlar

Yasemin Soysal Bu kişi benim

Feride Şahin Bu kişi benim

Sevda Menevşe Bu kişi benim

Yayımlanma Tarihi 26 Mayıs 2009
Gönderilme Tarihi 26 Mayıs 2009
Yayımlandığı Sayı Yıl 2008 Cilt: 15 Sayı: 3

Kaynak Göster

Vancouver Soysal Y, Şahin F, Menevşe S. Insan lenfosit hücre kültüründe melatonin ve b-karotenin mitomisin C ile indüklenmiş kardeş kromatid deşişimi sıklışı üzerine in vitro etkileri. SDÜ Tıp Fak Derg. 2009;15(3):23-9.

                                                                                         14791


Süleyman Demirel Üniversitesi Tıp Fakültesi Dergisi/Medical Journal of Süleyman Demirel University is licensed under Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International.