title: Exome-wide mutation profile in benzo[a]pyrene-derived post-stasis and immortal human mammary epithelial cells.
abstract: Genetic mutations are known to drive cancer progression and certain tumors have mutation signatures that reflect exposures to environmental carcinogens. Benzo[a]pyrene (BaP) has a known mutation signature and has proven capable of inducing changes to DNA sequence that drives normal pre-stasis human mammary epithelial cells (HMEC) past a first tumor suppressor barrier (stasis) and toward immortality. We analyzed normal, pre-stasis HMEC, three independent BaP-derived post-stasis HMEC strains (184Aa, 184Be, 184Ce) and two of their immortal derivatives(184A1 and 184BE1) by whole exome sequencing. The independent post-stasis strains exhibited between 93 and 233 BaP-induced mutations in exons. Seventy percent of the mutations were C:G>A:T transversions, consistent with the known mutation spectrum of BaP. Mutations predicted to impact protein function occurred in several known and putative cancer drivers including p16, PLCG1, MED12, TAF1 in 184Aa; PIK3CG, HSP90AB1, WHSC1L1, LCP1 in 184Be and FANCA, LPP in 184Ce. Biological processes that typically harbor cancer driver mutations such as cell cycle, regulation of cell death and proliferation, RNA processing, chromatin modification and DNA repair were found to have mutations predicted to impact function in each of the post-stasis strains. Spontaneously immortalized HMEC lines derived from two of the BaP-derived post-stasis strains shared greater than 95% of their BaP-induced mutations with their precursor cells. These immortal HMEC had 10 or fewer additional point mutations relative to their post-stasis precursors, but acquired chromosomal anomalies during immortalization that arose independent of BaP. The results of this study indicate that acute exposures of HMEC to high dose BaP recapitulate mutation patterns of human tumors and can induce mutations in a number of cancer driver genes.
authors.affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, AZ, 85724, USA.
authors.forename: Paul L
authors.initials: PL
authors.lastname: Severson
authors.affiliation: University of Arizona Cancer Center, Tucson, AZ, 85724, USA.
authors.forename: Lukas
authors.initials: L
authors.lastname: Vrba
authors.affiliation: University of Arizona Cancer Center, Tucson, AZ, 85724, USA; Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
authors.forename: Martha R
authors.initials: MR
authors.lastname: Stampfer
authors.affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, AZ, 85724, USA; University of Arizona Cancer Center, Tucson, AZ, 85724, USA. Electronic address: BFutscher@uacc.arizona.edu.
authors.forename: Bernard W
authors.initials: BW
authors.lastname: Futscher
mesh_terms: D001564:Benzo(a)pyrene / Q000633:toxicity*; D002274:Carcinogens, Environmental; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D004847:Epithelial Cells / Q000187:drug effects; D059472:Exome / Q000187:drug effects*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D052138:Genes, Neoplasm; D006801:Humans; D042361:Mammary Glands, Human / Q000166:cytology / Q000187:drug effects*; D009154:Mutation; D009369:Neoplasms / Q000139:chemically induced / Q000235:genetics; D017422:Sequence Analysis, DNA; D055815:Young Adult
chemical_list: D002274:Carcinogens, Environmental; D001564:Benzo(a)pyrene
keywords: Benzo[a]pyrene; Carcinogenesis; HMEC; p16