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Published on: February 1, 2011
[Genome DNA hypomethylation in HaCaT cells after short exposure to SiO2 nanoparticles]
Chunmei Gong1, Linqing Yang, Gonghua Tao
1Shenzhen Center for Chronic Disease Control, Shenzhen 518020, China. spring417@126.com
Objective:
To observe the effect of SiO2 nanoparticles on genome DNA methylation profile in cultured cells.
Methods:
HaCaT cells were treated with nm-SiO2 at 2.5, 5 and 10 microg/ml and micro-SiO2 at 10 microg/ml for 24h and DAC treatment was given at 10 microg/ml group for 48h. The mC/(mC + C) percent was quantified by high performance capillary electrophoresis (HPCE) assay, and the expression level of mRNA and protein was detected by Real-time Q-PCR and westernblot assay. The activity of DNMTs was determined by DNA Methyltransferase Activity/Inhibition Assay Kit.
Results:
HPCE assay showed that nm-SiO2-treated cells were decreased in some degree. An average proportion of methylated mC/ (mC + C) was 4.82% in control, 2.7% in 2.5 microg/ml and 2.17% in 10 microg/ml groups, while 3.1% in micro-SiO2 groups, which got the consistent downtrend of genome methylation level during increasing nm-SiO2 dose nanoparticles. The mRNA expression level for DNMT1 decreased gradually with increased dose of nm-SiO2 nanoparticles. The alterations at protein level were similar to those at the mRNA level.
Conclusion:
Genomic DNA methylation levels were decreased in HaCaT cells after short-term exposure to SiO2 nanoparticles.
