Year 2020, Volume 23 , Issue 1, Pages 247 - 258 2020-02-28

Evaluation of Antropogenic Aquatic Toxicity by and Alternative and New Approach; using PLHC-1 and RTG-2 cell lines and CYP1A1 Biomarker
Antropojenik Kaynaklı Sucul Toksisitenin Belirlenmesinde Alternatif ve Yeni Bir Yaklaşım Olarak PLHC-1 ve RTG-2 Hücre Hatlarının Kullanılması ve CYP1A1 Biyobelirteci ile Birlikte Değerlendirilmesi

Begüm YURDAKÖK DİKMEN [1] , Farah Gönül AYDIN [2] , Hidayet TUTUN [3] , Sedat SEVİN [4]


Permanent organic pollutants (POPs) are chemicals that can remain intact in nature for a long time and affect the health of humans, animals and the environment. In this study, cytotoxicity and 7-Ethoxyresorufin O-deethylase (EROD) activity of water (6 stations) and sediment (12 stations) samples collected from Ankara Stream for 12 months were evaluated on fish gonadal (RTG-2) and fish hepatocellular carcinoma (PLHC-1) cell lines. Cytotoxic activity were found to vary according to the sampling region and period. This study, provide important information for the methodology used in the detection of environmental contaminants and evaluation of pollution from environmental samples using sensitive, cheap, fast and trustable method by cytotoxicity and EROD activity in fish cells.  It is recommended  that these results could be optimized and used by legal authorities for risk evaluation

Kalıcı organik kirleticiler (KOK), insan, hayvan ve çevre sağlığını olumsuz etkileyen doğada uzun süre bozulmadan kalabilen ve çoğunlukla yağ dokuda birikme özelliği olan kimyasallardır. Bu araştırmada, 12 ay boyunca Ankara Çayı’ndan toplanan su (6 istasyon) ve sediment (12 istasyon) örneklerinin balık gonadal (RTG-2) ve balık hepatoselüler karsinom (PLHC-1) hücre hatlarında sitotoksitesi ve 7-etoksirezorufin O-deetilaz (EROD) etkinliği araştırılmıştır. Sitotoksik etkinliğin numune alınan bölge ve döneme göre değişkenlik göstermiştir. Bu araştırma ile biyomarker olarak kullanılan CYP1A düzeyi ve sitotoksisite değerlendirmelerinin balık hücrelerinde, çevresel örnekler üzerinde çalışılması ile kirlilik durumu ve kirleticiler hakkında ucuz, hızlı, tekrar edilebilir ve güvenilir bir ön-değerlendirme olduğu; ilgili kurumlar tarafından risk haritalarının çıkarılması amacıyla kullanılabileceği önerilmektedir.

  • Anonim 2012. OSB’de Atık Su Sancısı. http://www.hurriyet.com.tr/osb-de-atik-su-sancisi-21957791 ( Erişim Tarihi: 18.04.2019.)
  • Atıcı T, Ahıska S 2005. Pollution and algae of Ankara stream. Gazi Üniversitesi Fen Bilimleri Dergisi 18(1): 51-59.
  • Babin MM, Tarazona JV 2005. In vitro toxicity of selected pesticides on RTG-2 and RTL-W1 fish cell lines. Environ Polut 135(2):267-274.
  • Baksi SM, Frazier JM 1990. Isolated fish hepatocytes—model systems for toxicology research. Aquat Toxicol 16(4):229–256
  • Barile FA 1994. Introduction to In Vitro Cytotoxicology: Mechanisms and Methods. New York: CRC Press, p.: 33-43.
  • Behrens A, Schirmer K, Bols NC, Segner H 2001. Polycyclic aromatic hydrocarbons as inducers of cytochrome P4501A enzyme activity in the rainbow trout liver cell line, RTL-W1 and in primary cultures of rainbow trout hepatocytes. Environ Toxicol Chem 20(3) :632–643.
  • Bols NC, Dayeh VR, Lee LEJ, Schirme K 2005. Use of fish cell lines in the toxicology and ecotoxicology of fish. Piscine cell lines in environmental toxicology. Biochemistry and Molecular Biology of Fishes 6: 43-84.
  • Boyuneğmez T 2004. Biochemical monitoring of toxic and carcinogenic organic pollutants along the Izmir bay after the great canal project and possible health effects. ODTÜ Fen Bilimleri Enstitüsü, Biyokimya Anabilim Dalı, Yüksek Lisans Tezi, 121 syf.
  • Bradford MM 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248–254.
  • Bruschweiler BJ, Wurgler F, Fent K 1996 An ELISA for the determination of cytochrome P4501A in fish cell cultures. Environ Toxicol Chem 15:592-596
  • Butler M 2004. Animal Cell Culture and Technology, Taylor & Francis, New York p.: 32-33.
  • Clemons JH, Dixon DG, Bols NC 1997. Derivation of 2,3,7,8,-TCDD toxic equivalency factors (TEFs) for selected dioxins, furans and PCBs with rainbow trout and rat liver cell lines and the infuence of exposure time. Chemosphere 34 (5-7):1105-1119.
  • Cravedi JP, Paris A, Monod G, Devaux A, Flouriot G, Valotaire Y 1996. Maintenance of cytochrome P450 content and phase I and phase II enzyme activities in trout hepatocytes cultured as spheroidal aggregates. Comp Biochem Physiol C 113(2):241–246.
  • Davoren M, Shúilleabháin SN, Hartl MGJ, Sheehan D, O'brien NM, O'halloran J, Van Pelt FN, Mothersill C 2005. Assessing the potential of fish cell lines as tools for the cytotoxicity testing of estuarine sediment aqueous elutriates. Toxicol In Vitro 19(3): 421-31.
  • Devaux A, Pesonen M, Monod G, Andersson TB 1992. Gluco-corticoid- mediated potentiation on P450 induction in primary culture of rainbow trout hepatocytes. Biochem Pharmacol 43(4):898-901
  • Dubois M, Plaisance H, Thome JP, Kremers P 1996. Hierarchical cluster analysis of environmental pollutants through P450 induction in cultured hepatic cells. Ecotoxicol. Environ. Saf 34(3): 205-215.
  • Eichbaum K, Brinkmann M, Buchinger S, Reifferscheid G, Hecker M, Giesy JP, Engwall M, Van Bavel B, Hollert H 2014. In vitro bioassays for detecting dioxin-like activity—Application potentials and limits of detection, a review. Science of the Total Environment 487: 37-48.
  • Fent K 2007. Permanent Fish Cell Cultures as Important Tools in Ecotoxicology. ALTEX 24(8): 26-28.
  • Fotakis G, Tımbrell JA 2006. In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicology Letters 160 (2): 171–177.
  • García-Tavera JL, Valdés-Lozano D, Poblete-Naredo I, Albores-Medina A, Zapata-Pérez O 2013. Bile benzo[a]pyrene concentration and hepatic CYP1A induction in hypoxic adult tilapia (Oreochromis niloticus). Chemosphere 92 (1):16–23.
  • Gedik K, Imamoglu I 2011. Assessment of temporal variation and sources of PCBs in the sediments of Mediterranean Sea, Mersin Bay, Turkey. Marine Pollution Bulletin 62(1): 173–177.
  • Gedik K, Imamoglu I. 2013. Levels, Distribution, and Sources of Polychlorinated Biphenyls in Sediments of Lake Eymir, Turkey. Arch Environ Contam Toxicol 65(2): 203-211.
  • Gökalp Z, Tas I, Ozkay F, Akgül S 2011. Wastewater use in irrigation-Ankara case study. African Journal of Agricultural Research 67(7): 1807-1812.
  • Hahn ME, Woodward BL, Stegeman JJ, Kennedy SW 1996. Rapid assessment of induced cytochrome P4501A protein and catalytic activity in fish hepatoma cells grown in multiwell plate: response to TCDD, TCDF and two planar PCBs. Environ Toxicol Chem 15 (4):582-591
  • Huuskonen SE, Tuvikene A, Trapido M, Fent K, Hahn ME 2000. Cytochrome P4501A induction and porphyrin accumulation in PLHC-1 fish cells exposed to sediment and oil shale extracts. Archives of environmental contamination and toxicology 38(1): 59-69.
  • Isomaa B, Lilius H 1995. The urgent need for in vitro tests in ecotoxicology. Toxic In Vitro 9(6):821–825
  • Karakoç G, Erkoç FU, Katırcıoglu H 2003. Water quality and impacts of pollution sources for Eymir and Mogan Lakes (Turkey). Environ Int 29(1): 21-27.
  • Kazancı N, Girgin S 1998. Distribution of Oligocheta species as bioindicators of organic pollution in Ankara Stream and their use in biomonitoring. Turkish Journal of Zoology 22: 83-87.
  • Machala M, Nezveda K, Petrivalsky M, Jarosova A, Piacka V, Svobodova Z 1997. Monooxygenase activities in carp as biochemical markers of pollution by polycyclic and polyhalogenated aromatic hydrocarbons: choice of substrates and effects of temperature, gender and capture stress. Aquatic Toxicology 37: 113-123.
  • Machala M, Vondracek J, Blaha L, Ciganek M, Neca J 2001. Aryl hydrocarbon receptor mediated activity of mutagenic polycyclic aromatic hydrocarbons determined using in vitro reporter gene assay. Mutation Research 497: 49-62.
  • Mosmann T 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63.
  • Özmen M, Gungordu A, Kucukbay FZ, Guler RE 2006. Monitoring the effects of water pollution on Cyprinus carpio in Karakaya Dam Lake, Turkey. Ecotoxicology 15: 157-69.
  • Pesonen M, Goksoyr A, Andersson T 1992. Expression of P4501A1 in a primary culture of rainbow trout hepatocytes exposed to bnapthoflavone or 2,3,7,8-tetrachlorodibenzo-p-dioxin. Arch Biochem Biophys 292(1):228–233
  • Repetto G, Del Peso A, Zurıta JL 2008. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nature Protocols, 3(7): 1125-1131.
  • Scholz S, Segner H 1999. Induction of CYP1A in primary cultures of rainbow trout (Oncorhynchus mykiss) liver cells: Concentration–response relationships of four model substances. Ecotoxicology and environmental safety 43(3): 252-260.
  • Segner H, Lenz D, Hanke W, Schuurmann G 1994. Cytotoxicity of metals toward rainbow trout R1 cell line. Environ. Toxicol. Water Quality 9(4): 273–279.
  • Sijm DTHM, Schaap G, Opperhuizen A 1993. The elect of the biotransformation inhibitor piperonylbutoxide on the bioconcentration of 2,8-dichlorodibenzo-P-dioxin and pentachlorobenzene in goldfsh. Aquat. toxicol 27 (3-4): 345-360.
  • Siroka Z, Drasticova J 2004. Biochemical Markers of Aquatic Environment Contamination - Cytochrome P450 in Fish. A Review. Acta Veterinaria Brno 73 (1): 123-132.
  • Suares-Rocha P, Braunbeck T, De Angelis DDF, Marin-Morales MA 2015. Assessment of cytotoxicity and AhR-mediated toxicity in tropical fresh water sediments under the influence of an oil refinery. Environmental Science and Pollution Research 22(16), 12566-12575.
  • Timur UP, Barış ME 2014. Greenway Planning Based on River for Sustainable Cities: An Example of Ankara City. International Journal of Natural and Engineering Sciences 8(2): 35-41.
  • Traven L, Zaja R, Loncar J, Smital T, Mićović V 2008. CYP1A induction potential and the concentration of priority pollutants in marine sediment samples--in vitro evaluation using the PLHC-1 fish hepatoma cell line. Toxicol In Vitro 22 (6): 1648-56.
  • Ulukaya E, Ozdikicioglu F, Oral AY, Demirci M 2008. The MTT assay yields a relatively lower result of growth inhibition than the ATP assay depending on the chemotherapeutic drugs tested. Toxicology in vitro 22(1): 232–239.
  • UNEP 1999. United Nations Environmental Program. Guidelines for the Identification of PCBs and Materials Containing PCBs.: 40
  • Uno T, Ishizuka M, Itakura T 2012. Cytochrome P450 (CYP) in fish. Environ Toxicol Pharmacol 34(1):1–13.
  • White RD, Shea D, Solow AR, Stegeman JJ 1997. Induction and post-transcriptional suppression of hepatic cytochrome P450 1A1 by 3,3′,4,4′-tetrachlorobiphenyl. Biochem Pharmacol 53(7): 1029–1040.
Primary Language tr
Subjects Science
Journal Section RESEARCH ARTICLE
Authors

Orcid: 0000-0002-0385-3602
Author: Begüm YURDAKÖK DİKMEN
Institution: ANKARA ÜNİVERSİTESİ, VETERİNER FAKÜLTESİ, KLİNİK ÖNCESİ BİLİMLERİ BÖLÜMÜ, VETERİNERLİK FARMAKOLOJİ VE TOKSİKOLOJİSİ ANABİLİM DALI
Country: Turkey


Orcid: 0000-0002-0068-2078
Author: Farah Gönül AYDIN (Primary Author)
Institution: ANKARA ÜNİVERSİTESİ, VETERİNER FAKÜLTESİ, KLİNİK ÖNCESİ BİLİMLERİ BÖLÜMÜ, VETERİNERLİK FARMAKOLOJİ VE TOKSİKOLOJİSİ ANABİLİM DALI
Country: Turkey


Orcid: 0000-0001-9512-8637
Author: Hidayet TUTUN
Institution: MEHMET AKİF ERSOY ÜNİVERSİTESİ, VETERİNER FAKÜLTESİ, KLİNİK ÖNCESİ BİLİMLER BÖLÜMÜ, VETERİNERLİK FARMAKOLOJİ VE TOKSİKOLOJİSİ ANABİLİM DALI
Country: Turkey


Orcid: 0000-0003-0475-9092
Author: Sedat SEVİN
Institution: ANKARA ÜNİVERSİTESİ, VETERİNER FAKÜLTESİ, KLİNİK ÖNCESİ BİLİMLERİ BÖLÜMÜ, VETERİNERLİK FARMAKOLOJİ VE TOKSİKOLOJİSİ ANABİLİM DALI
Country: Turkey


Supporting Institution Ankara Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü
Project Number 13 B 3338010
Thanks Bu çalışma, Ankara Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü’nce 13B3338010 numaralı proje ile desteklenmiştir.
Dates

Application Date : April 22, 2019
Acceptance Date : August 5, 2019
Publication Date : February 28, 2020

APA YURDAKÖK DİKMEN, B , AYDIN, F , TUTUN, H , SEVİN, S . (2020). Antropojenik Kaynaklı Sucul Toksisitenin Belirlenmesinde Alternatif ve Yeni Bir Yaklaşım Olarak PLHC-1 ve RTG-2 Hücre Hatlarının Kullanılması ve CYP1A1 Biyobelirteci ile Birlikte Değerlendirilmesi. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi , 23 (1) , 247-258 . DOI: 10.18016/ksutarimdoga.vi.556762