N-Acetyltransferase 1 Polymorphism and Breast Cancer Risk
Abstract
N-acetyltransferase 1 (NAT1) catalyzes N-acetylation of aryl amine carcinogens resulting in their activation or inactivation. NAT1*10, NAT1*11 and NAT1*14, common variant alleles have been epidemiologically associated with increased risk for numerous cancers including breast. NAT1 is also upregulated in breast cancer. We employed a novel approach to study functional differences caused by NAT1*10 , NAT1*11, and NAT1*14 polymorphisms by using constructs that mimic complete human mRNAs. Plasmid constructs of NAT1*10 and NAT1*4 contained full length human mRNAs including either the NATa (alternative promoter) or NATb (major promoter) 5?-UTR, the ORF, and 885 base pairs of the 3'UTR region. Following transient or stable transfection into Chinese hamster ovary cells, NAT1-catalyzed N-acetylation of p-aminobenzoic acid was measured by HPLC and NAT1 protein expression was measured by Western blot. mRNA levels were studied using RT-PCR and polyA patterns by RNase Protection. No differences were observed in acetylation activity, protein levels, mRNA, or polyA patterns between NAT1*10, NAT1*11 and NAT1*4, but significant differences were seen between NAT1*14 and NAT1*4. Significant differences were also seen between all constructs containing the NATb 5' UTR compared to those containing the NATa 5' UTR.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2009
- Accession Number
- ADA516365
Entities
People
- Lori M. Millner
Organizations
- University of Louisville