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J. Biol. Chem., Vol. 261, Issue 4, 1594-1598, 02, 1986
TL Kautiainen and PA Jones
The levels of DNA methyltransferase in nuclei from 9 tumorigenic and 9
nontumorigenic cell lines were examined. In all but 2 cases, the
extractable methyltransferase activity was 4-3000-fold higher in
tumorigenic than in nontumorigenic cells. Tumorigenic and nontumorigenic
cells from four species were grown in the presence of various
concentrations (10(-8)-10(-6) M) of an inhibitor of the methylase enzyme,
5-aza-2'-deoxycytidine (5-aza-dCyd). The reduction of 5-methylcytosine
content in newly replicated DNA in the presence of 5- aza-dCyd was used to
determine the relative methylase activity in each cell line. In all 4
cases, tumorigenic cells required larger doses of drug to inhibit DNA
methylation to the same extent as their nontumorigenic counterparts. The
relative rates of incorporation of [3H]5-aza-dCyd were determined for each
cell line, and tumorigenic cells were shown to incorporate equal or greater
amounts of 5-aza-dCyd into DNA compared to nontumorigenic cells. These
results showed that the differences in the inhibition of DNA methylation in
response to 5- aza-dCyd were not due to differences in the ability of these
cells to incorporate the drug. Thus, it was demonstrated by two independent
methods that tumorigenic cells contained higher levels of methylating
capacity than nontumorigenic cells. This overabundance of methyltransferase
may alter DNA methylation patterns and affect phenotypic stability.
DNA methyltransferase levels in tumorigenic and nontumorigenic cells in culture
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