The investigation of mutagenic and carcinogenic effects by the ames test

Year 2013, Volume: 3 Issue: 2, 75 - 82, 30.01.2014

Selin Oğuz Gülden Z. Omurtag , Feyza Arıcıoğlu Semra Şardaş

Abstract

Aim: Research has shown that the increased use of chemicals which are offered to consumers, sometimes without sufficient testing, has impacted human health and natural resources negatively. For this reason, many researchers have developed cost-effective test systems which can examine the carcinogenic and mutagenic activity of chemicals in a short time. Ames is an example of such a test system which can detect mutations at cellular level. Also the Ames test is accepted as a reliable method for specifically testing the toxic and mutagenic-carcinogenic effects of raw materials to be used in pharmaceuticals.

Material and Methods: In this study, the mutagenic and carcinogenic effect of methylphenidate, that is used for the treatment of Attention-Deficit Hyperactivity Disorder, was investigated by the Ames Salmonella/microsome test in the absence and presence of metabolic activation. The Ames Salmonella/microsome test can play an important role to identify the mutagenic effects of carcinogenic compounds and is used for the research and development of drugs. TA 98 and TA 100 strains were used in experiments. TA 98 is designed for frame-shift mutagens and TA 100 is designed for base-pair mutagens. The mutagenic activity was screened in two groups with or without S9 metabolic activation. The results were evaluated and the mean values were compared with positive and negative controls.

Results: According to the results of experiments conducted with TA98 and TA100, revertant colony count was not found higher than those spontaneous revertant colony count among all of the concentrations and as well as under the conditions of with and without S9 (p>0.05).

Conclusion: In conclusion, it was shown that methylphenidate was non-mutagenic for TA 98 and TA 100 with or without S9 metabolic activation.

Key words: Ames test, carcinogenicity, methylphenidate, S9 fraction, mutagenicity

Keywords

Ames test, carcinogenicity, methylphenidate, S9 fraction, mutagenicity

Mutajenik-karsinojenik etkinin ames testi ile araştırılması

Abstract

Amaç: Kimyasalların insan hayatında daha fazla yer alması ve buna bağlı olarak kimyasal maddelerin gerekli güvenlik testlerinden geçmeden kullanılmaya başlanmasıyla birlikte kimyasalların insan sağlığı ve doğal kaynaklar üzerine olumsuz etkileri ortaya çıkmıştır. Bu sebeple birçok araştırmacı, kimyasalların karsinojenik ve mutajenik aktivitelerini inceleyip kısa zamanda sonuç veren düşük maliyetli mutajenite test sistemleri geliştirmişlerdir. Kimyasallar tarafından meydana gelen mutasyonların hücre düzeyinde belirlenmesi amacıyla kullanılan kısa zamanlı test sistemlerinden biri de Ames testidir. Ames testi, özellikle ilaç ham maddesi olarak kullanılmak istenen test maddelerinin toksik, mutajenik-karsinojenik etkilerini incelemek için güvenilir yöntemler arasında kabul edilmektedir.

Gereç ve Yöntem: Bu çalışma ile ilaçların geliştirilmesinde ve karsinojen maddelerin mutajenitelerinin saptanmasında kullanılan önemli bir mutajenite testi olarak kabul edilen Ames Salmonella/mikrozom Testi uygulanarak dikkat eksikliği ve hiperaktivite bozukluğunda kullanılan ilaç ham maddesi metilfenidatın mutajenik ve karsinojenik etkisi araştırılmıştır. Yöntemde Salmonella typhimurium TA 100 ve TA 98 suşları ile çalışılmıştır.

Bulgular: TA 100 baz çifti değişimine, TA 98 ise çerçeve kaymasına yol açan mutajenlerin belirlenmesi için kullanılmıştır. Deneyler S9’lu ve S9’suz olmak üzere iki grup halinde gerçekleştirilmiştir. Test sonuçlarının ortalaması alınarak, pozitif ve negatif kontrol grupları ile karşılaştırılmış ve değerlendirilmiştir.

Sonuç: Çalışmamızda, metilfenidatın TA 100 ve TA 98 suşları ile S9 varlığında ve yokluğunda mutajenik aktiviteye sahip olmadığı gösterilmiştir.

Anahtar Kelimeler : Ames testi, karsinojenite, metilfenidat, mutajenite, S9 fraksiyonu

Keywords

Ames testi, karsinojenite, metilfenidat, mutajenite, S9 fraksiyonu

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