Abstract
<h2>Abstract</h2><h3>Background/Aims</h3> Examine the mitoinhibitory effect of the liver tumor promoter 2-acetylaminofluorene (2-AAF) in vivo, with focus on the proteins regulating G1- and S progression. <h3>Methods</h3> cdk 2 kinase assay to examine histone H1 phosphorylation. cdk 4 kinase assay to examine whether active cdk 4/cyclin D complexes, capable of phosphorylating Rb, are formed. Western blot to monitor protein expression. <h3>Results</h3> cdk 4 kinase-mediated Rb phosphorylation was increased by AAF treatment. Nuclear expression of the transcription factors E2F-1 and E2F-3 was downregulated, while E2F-4 was decreased. 2-AAF treatment also markedly reduced cdk 2 kinase activity/histone H1 phosphorylation during G1/S-transition. Western blot showed loss of nuclear as well as cytoplasmic cyclin A and cyclin B protein after 2-AAF treatment, while the Rb protein level was markedly increased during G1. The cell cycle dependent elevation of nuclear p107 protein, seen in control livers, was not observed in AAF-treated animals. <h3>Conclusions</h3> Effects of 2-AAF; Very low cdk 2 kinase activity that could possibly block G1/S-transition; increased pRb level together with diminished levels of transcription factors E2F-1 and -3, that could be responsible for reducing the expression of E2F target genes such as cyclin A and E2F-1.
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