Journal Article

Modulation by budesonide of a CpG endonuclease in mouse lung tumors

Long Li, Lianhui Tao, Ronald A. Lubet, Vernon E. Steele and Michael A. Pereira

in Carcinogenesis

Volume 28, issue 7, pages 1499-1503
Published in print July 2007 | ISSN: 0143-3334
Published online March 2007 | e-ISSN: 1460-2180 | DOI:
Modulation by budesonide of a CpG endonuclease in mouse lung tumors

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CpG endonuclease activity was identified in nuclear extracts obtained from mouse lung tumors. Enzyme activity was determined using a 333 bp polymerase chain reaction product of the estrogen receptor-α gene that contained either radiolabeled cytosine or tritium-labeled methyl groups at CpG sites. Activity was measured as the release of radioactivity from the substrate. The product of the nuclease activity was identified by high pressure liquid chromatography (HPLC) as either 5-methyl-2′-deoxycytidine when the CpG sites in the substrate were methylated or 2′-deoxycytidine when the CpG sites were not methylated. The CpG endonuclease activity was dependent on nuclear protein and temperature, had a proclivity for double-stranded over single-stranded DNA and was inhibited by ethylenediaminetetraacetic acid or 2-mercaptoethanol. Strain A/J mouse lung tumors induced by vinyl carbamate had a greater level of CpG endonuclease activity than non-involved lung tissue. Budesonide, a potent chemopreventive agent in mouse lung, not only prevented an increase in CpG endonuclease activity in lung tumors but, when administered to mice with established tumors, also decreased the level of endonuclease activity in the tumors. The effect of budesonide on CpG endonuclease activity in lung tumors was inversely related to its published effect on DNA methylation in mouse lung tumors, i.e. the drug decreased CpG endonuclease activity and increased the methylation of DNA. The increased CpG endonuclease activity in mouse lung tumors and its inhibition by budesonide would suggest this endonuclease as a potential molecular target for chemoprevention.

Journal Article.  4187 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

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