Study on GMA-DNA adducts.
OBJECTIVE
DNA modification fixed as mutations in the cells may be an essential factor in the initiation step of chemical carcinogenesis. In order to explore the mechanism of gene mutation and cell transformation induced by glycidyl methacrylate (GMA), the current test studied the characteristics of GMA-DNA adducts formation in vitro.


METHODS
In vitro test, dAMP, dCMP, dGMP, dTMP and calf thymus DNA were allowed to react with GMA (Glycidyl Methacrylate). After the reaction, the mixtures were detected by UV and subjected to reversed-phase HPLC on ultrasphere ODS reversed-phase column, the reaction products were eluted with a linear gradients of methanol (solvent A) and 10 mmol/L ammonium formate, pH 5.0 (solvent B). The synthesized adducts were then characterized by UV spectroscopy in acid (pH 1.0), neutral (pH 7.2), alkaline (pH 11.0) and by mass spectroscopy.


RESULTS
The results showed that GMA could bind with dAMP, dCMP, dGMP and calf thymus DNA by covalent bond, and the binding sites were specific (N6 of adenine, N3 of cytosine). Meanwhile, a main GMA-DNA adduct in the reaction of GMA with calf thymus DNA was confirmed as N3-methacrylate-2-hydroxypropyl-dCMP.


CONCLUSIONS
GMA can react with DNA and/or deoxynucleotide monophosphate and generate some adducts such as N6-methacrylate-2-hydroxypropyl-dAMP and N3-methacrylate-2-hydroxypropyl-dCMP, ets. Formation of GMA-DNA adducts is an important molecular event in gene mutation and cell transformation induced by GMA.
Fude
F
F
Institute of Basic Medical Sciences, CAMS & PUMC, Beijing 100005.
Jin
Z
Z
Haixin
L
L
Mingjia
T
T
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism*; D018736:DNA Adducts / Q000096:biosynthesis*; D003843:Deoxycytidine Monophosphate / Q000378:metabolism; D003848:Deoxyguanine Nucleotides / Q000378:metabolism; D004852:Epoxy Compounds / Q000494:pharmacology*; D066298:In Vitro Techniques; D008689:Methacrylates / Q000494:pharmacology*; D013056:Spectrophotometry, Ultraviolet; D013938:Thymidine Monophosphate / Q000378:metabolism
D018736:DNA Adducts; D003848:Deoxyguanine Nucleotides; D004852:Epoxy Compounds; D008689:Methacrylates; D003843:Deoxycytidine Monophosphate; D013938:Thymidine Monophosphate; C007257:2'-deoxyguanosine 5'-phosphate; D004247:DNA; C007870:glycidyl methacrylate