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Analisis Kadar O6-(H3) Metilguanin-DNA pada Kultur Hepatosit Tikus Menggunakan Liquid Scintillation Counter Setelah Pemberian (-)-Epigalokatekin Galat dari Teh Hijau

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Background.Alkylation of DNA can occur at various site, however lkylation of the O6-position ofguanine has the strongest mutagenic potential. O6
-Methylguanine is a toxic and mutagenic lesionwhich is formed in cellular DNA by alkylating agents. Alkyl group from carbanium ion can attack DNA
forms alkylated DNA especially at O6 position of guanine. O6-ethylguanine mispairs with thymine during replication, and if the adduct is not emoved, this results in conversion from a guanine-cytosine pair to an adenine-thymine pair. Deficient repair of O6-methylguanine has been suggested to be a contributory factor in the etiology of some diseases such as diabetes and cancer. This study aimed to determine - ethylguanine DNA concentration induced by N-[H3]methyl-N-nitrosourea ([H3]MNU)in rat liver cell culture with and without (-)-epigalocatechin gallate exposure.
Methods.Primary rat liver cell culture was divided into 10 groups. Five groups for [H3] MNU 32 μMexposure, and 5 groups remain for 48 μM [H3]MNU exposure. Each group of [H3]MNU exposure got EGCG 0 (as control), 8, 17, 33 and 67 ppm. The DNA was isolated, and O6-[H3]methylguanine DNA
separated by high performance liquid chromatography (HPLC). Concentration of O6-[H3]methylguanine DNA was measured by Liquid Scintillation Counter.
Results.O6-Methylguanine DNA concentration decreased significantly in liver cell culture groups with EGCG. O6-[H3 ]Methylguanine DNA concentration in control groups for [H3]MNU 32 μM and 48 μM were 29.89 ± 2.01 and 32.00 ± 1.67 fmol/μg DNA, respectively, while with EGCG 67 ppm were 1.87 ± 0.94 and 2.38 ± 0.36 fmol/μg DNA.
Conclusion.O6-methylguanine DNA concentration in liver rat culture induced by MNU decreases significantly after EGCG exposure. These findings support the conclusion that EGCG plays a key role in
suppressing and inhibiting cancer development.


-[H
3
]methyl-
N
-nitrosou

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Call Number

Jurnal Kedokteran Indonesia,2(1) 2011:5-10

Publisher

Surakarta : Fk. Universitas Sebelas Maret., 2011

Collation

5p.

Language

ISBN/ISSN

2086-5171

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Edition

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Subject(s)

rats
tea
HEPATOCYTES

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