18 research outputs found
Epigenetic Factors in Cancer Risk: Effect of Chemical Carcinogens on Global DNA Methylation Pattern in Human TK6 Cells
In the current study, we assessed the global DNA methylation changes in human lymphoblastoid (TK6) cells in vitro in response to 5 direct and 10 indirect-acting genotoxic agents. TK6 cells were exposed to the selected agents for 24 h in the presence and/or absence of S9 metabolic mix. Liquid chromatography-mass spectrometry was used for quantitative profiling of 5-methyl-2′-deoxycytidine. The effect of exposure on 5-methyl-2′-deoxycytidine between control and exposed cultures was assessed by applying the marginal model with correlated residuals on % global DNA methylation data. We reported the induction of global DNA hypomethylation in TK6 cells in response to S9 metabolic mix, under the current experimental settings. Benzene, hydroquinone, styrene, carbon tetrachloride and trichloroethylene induced global DNA hypomethylation in TK6 cells. Furthermore, we showed that dose did not have an effect on global DNA methylation in TK6 cells. In conclusion we report changes in global DNA methylation as an early event in response to agents traditionally considered as genotoxic
Disruption of an AP-2 alpha binding site in an IRF6 enhancer is associated with cleft lip
Previously we have shown that nonsyndromic cleft lip with or
without cleft palate (NSCL/P)1 is strongly associated with
SNPs in IRF6 (interferon regulatory factor 6)2. Here, we use
multispecies sequence comparisons to identify a common SNP
(rs642961, G4A) in a newly identified IRF6 enhancer. The
A allele is significantly overtransmitted (P ¼ 1 1011) in
families with NSCL/P, in particular those with cleft lip but not
cleft palate. Further, there is a dosage effect of the A allele,
with a relative risk for cleft lip of 1.68 for the AG genotype
and 2.40 for the AA genotype. EMSA and ChIP assays
demonstrate that the risk allele disrupts the binding site of
transcription factor AP-2a and expression analysis in the
mouse localizes the enhancer activity to craniofacial and limb
structures. Our findings place IRF6 and AP-2a in the same
developmental pathway and identify a high-frequency variant
in a regulatory element contributing substantially to a
common, complex disorder