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J. Biol. Chem., Vol. 279, Issue 33, 34138-34149, August 13, 2004

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Metallothionein-III Prevents -Ray-induced 8-Oxoguanine Accumulation in Normal and hOGG1-depleted Cells^*

Hye Gwang Jeong, Cha-Kyung Youn, Hyun-Ju Cho, Soo-Hyun Kim, Mi-Hwa Kim, Hong-Beum Kim, In-Youb Chang, Yun-Sil Lee¶, Myung-Hee Chung||, and Ho Jin You¶**

From the Research Center for Proteineous Materials, Chosun University, 375 Seosuk-dong, Gwangju 501-759, Korea, the Department of Pharmacology, School of Medicine, Chosun University, 375 Seosuk-dong, Gwangju 501-759, Korea, the ^¶Laboratory of Radiation Effect, Korea Cancer Center Hospital, 215-4 Gongneung-Dong, Nowon-Ku, Seoul 139-706, Korea, and the ^||Department of Pharmacology, School of Medicine, Seoul National University, 28 Yongon-dong, Seoul 110-799, Korea

Metallothioneins (MT) play an important biological role in preventing oxidative damage to cells. We have previously demonstrated that the efficiency of the protective effect of MT-III against the DNA degradation from oxidative damage was much higher than that of MT-I/II. As an extension of the latter investigation, this study aimed to assess the ability of MT-III to suppress 8-oxoguanine (8-oxoG), which is one of the major base lesions formed after an oxidative attack to DNA and the mutant frequency of the HPRT gene in human fibroblast GM00637 cells upon exposure to -rays. We found that human MT-III expression decreased the level of 8-oxoG and mutation frequency in the -irradiated cells. Using an 8-oxoguanine DNA glycosylase (OGG1)-specific siRNAs, we also found that MT-III expression resulted in the suppression of the -radiation-induced 8-oxoG accumulation and mutation in the OGG1-depleted cells. Moreover, the down-regulation of MT in human neuroblastoma SKNSH cells induced by MT-specific siRNA led to a significant increase in the 8-oxoG level, after exposure to -irradiation. These results suggest that under the conditions of -ray oxidative stress, MT-III prevents the -radiation-induced 8-oxoG accumulation and mutation in normal and hOGG1-depleted cells, and this suppression might, at least in part, contribute to the anticarcinogenic and neuroprotective role of MT-III.

Received for publication, March 5, 2004 , and in revised form, June 8, 2004.

^* This work was supported by Grant M1-0311-00-0077 from the Molecular and Cellular BioDiscovery Research Program from the Ministry of Science and Technology and by the National Project of Nuclear R&D Program from Ministry of Science and Technology of Korea. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^** To whom correspondence should be addressed. Tel.: 82-62-230-6337; Fax: 82-62-233-3720; E-mail: hjyou{at}chosun.ac.kr.

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