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Genotyping of a p73 Dinucleotide Polymorphism in Human Cancer Cell Lines
Author(s) -
Cordova Ricardo,
Hampton Kaia,
Kim Dongwha,
Park Jong,
Carastro L Michael
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.576.7
Subject(s) - genotype , genotyping , microbiology and biotechnology , allele , promoter , genomic dna , exon , biology , gene isoform , gene , taqman , cell culture , genetics , tumor suppressor gene , cancer research , gene expression , carcinogenesis , polymerase chain reaction
The p73 gene is a member of the p53 tumor suppressor family. A p73 dinucleotide polymorphism (DNP) (rs1801173) is a G4C14‐to‐A4T14 linked pair of transitions located in exon 2 and lies between the P1 and P2 gene promoters. The P1 and P2 p73 gene promoters are the transcription initiation sites for mRNAs encoding TAp73 and DNp73 isoforms, respectively. Recently, we reported the p73 DNP allele was associated with (1) decreased risk [OR = 0.55, 95%CI =0.31‐0.99] for aggressive prostate cancer (PCa) and (2) increased TAp73/DNp73 protein isoform ratios in ten human cancer cell lines. We hypothesize the presence of the p73 DNP allele causes altered p73 promoter usage, ultimately resulting in increased TAp73/DNp73 protein isoform ratios, which could explain the observed decreased risk for PCa aggressiveness. Our ultimate goal is to assess the potential of p73 DNP as a biomarker for lower aggressive PCa risk. Therefore, our initial goal in this study was to determine the p73 DNP genotype in four human cancer cell lines (DU 145, MDA‐MB‐468, PC‐3 and SW48) not yet characterized for p73 DNP status. We previously characterized the p73 DNP genotype of three human cancer cell lines that we used as genotyping controls: Caco‐2 (homozygous polymorphic), NCI‐H1299 (homozygous wild type), and HepG2 (heterozygous). These seven cell lines were cultured and genomic DNA was isolated and quantitated. Cell line genomic DNA samples were used in TaqMan Real Time‐PCR allelic discrimination assays to determine the p73 DNP genotypes. Our data conclusively determined that DU 145, MDA‐MB‐468, and SW48 were p73 DNP homozygous wild type, and PC‐3 was p73 DNP heterozygous.