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Djoko Agus Purwanto
Departemen Ilmu Kefarmasian, Fakultas Farmasi Universitas Airlangga, Indonesia
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Journal : Jurnal Kedokteran Indonesia

 1 
Analisis Kadar O6 -[H3 ]Metilguanin-DNA pada Kultur Hepatosit Tikus Menggunakan Liquid Scintillation Counter Setelah Pemberian (-)-Epigalokatekin Galat dari Teh Hijau Purwanto, Djoko Agus
Jurnal Kedokteran Indonesia Vol 2, No 1 (2011)
Publisher : Jurnal Kedokteran Indonesia

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Abstract

Background: Alkylation of DNA can occur at various site, however alkylation 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 DNAforms alkylated DNA especially at O6 position of guanine. O6-methylguanine mispairs with thymineduring replication, and if the adduct is not removed, this results in conversion from a guanine-cytosinepair to an adenine-thymine pair. Deficient repair of O6-methylguanine has been suggested to be acontributory factor in the etiology of some diseases such as diabetes and cancer. This study aimed todetermine O6-methylguanine 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 gotEGCG 0 (as control), 8, 17, 33 and 67 ppm. The DNA was isolated, and O6-[H3]methylguanine DNAseparated by high performance liquid chromatography (HPLC). Concentration of O6-[H3]methylguanineDNA was measured by Liquid Scintillation Counter.Results: O6-Methylguanine DNA concentration decreased significantly in liver cell culture groups withEGCG. O6-[H3]Methylguanine DNA concentration in control groups for [H3]MNU 32 μM and 48 μMwere 29.89 ± 2.01 and 32.00 ± 1.67 fmol/μg DNA, respectively, while with EGCG 67 ppm were 1.87 ± 0.94and 2.38 ± 0.36 fmol/μg DNA.Conclusion: O6-methylguanine DNA concentration in liver rat culture induced by MNU decreasessignificantly after EGCG exposure. These findings support the conclusion that EGCG plays a key role insuppressing and inhibiting cancer development.Keywords:(-)-epigalocatechin gallate, N-[H3]methyl-N-nitrosourea, O6-methylguanine DNA, LiquidScintilation Counter
Co-Authors A Fahmy Arif Tsani A. Toto Purnomo Abby Rahmat Kamaruzzaman Achmad Fuad Hafid Achmad Toto Poernomo Achmad Toto Purnomo Adecha Ekalipta Primazafira Adi Utarini Afra, Fairuz Yaumil Agustina, Indri Alief Putriana Rahman Anggita Mirzautika Anita Natalia Suryawijaya Asri Darmawati Asri Darmawati Aty Widyawaruyanti Audrey Gracelia Riwu Augustien, Nora Budi Suharjo Budi Wikeko Christina Destri Corry, Maylisa Natalia Devi Eka Juniarti Dewi Isadiartuti Dewi Melani Hariyadi Edhi, Narendrani Sasmitaning Erwin Astha Triyono Esti Hendradi Fairuz Yaumil Afra Fauziyah, Riska Salfa Febri Annuryanti Guniarti Guniarti Guniarti Hadi Poerwono Hadi Suhardjono Hanifah Ridha Rabbani Hasan, Muhammad Amir Heny K Daryanto Herin Mawarti Hidayati, Agriana Rosmalina I Nyoman Wijaya Ilmi, Hilkatul Innisa, Deana Rohali Ira Widjiastuti Isnaeni Isnaeni Isnaeni Iswajuni, Iswajuni Joewono Soeroso Juni Ekowati Jusak Nugraha Jusak Nugraha Kun Ismiyatin Lahardo, Devanus M. Rasjad Indra Mangestuti Agil Marcellino Rudyanto Masruratos Zahroh Melanny Ika Sulistyowaty Mirzautika, Anggita Muhammad Amin Muslihatin, Wirda Noorma Rosita Nora Augustien Pribadi, Didik Huswo Purwaningsih - Purwitasari, Neny Retno Pudji Rahayu Retno Widyowati Riesta Primaharinastiti Riesta Primaharinastiti, Riesta Rufaidah Azzahrah Sabri, Mohamad Fazli Saribatiara, Saribatiara Setyabudi Setyabudi Soraya Aulia Sri Hartono Sri Iswati Suciati Suciati Sukardiman Susetyowati Susilowati Susilowati Sutini Tiara Sukmawati Toto Sudargo Tri Widiandani Tristiana Erawati Tutiek Purwanti Ujang Sumarwan Wibowo, Nadya Khadijah Wicaksana, Firman Widiwurjani Widji Soeratri Widji Soeratri Yayuk Hartriyanti Yudianingtyas, Deanur Yusniasari, Putri Antika Yusuf, Helmy