Mutagenicity of acrylamide and glycidamide in human TP53 knock-in (Hupki) mouse embryo fibroblasts

Hölzl-Armstrong, Lisa; Kucab, Jill E.; Moody, Sarah; Zwart, Edwin; Loutkotová, Lucie; Duffy, Veronica; Luijten, Mirjam; Costa, Gonçalo Gamboa da; Stratton, Michael R.; Phillips, David H.; Arlt, Volker M.

doi:10.1007/s00204-020-02878-0

Cited by 26 publications

(36 citation statements)

References 51 publications

(124 reference statements)

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“…A TP53-WT culture was included to demonstrate a typical response towards Nutlin-3a treatment, which is reflected by an induction of all three proteins. In TP53- Gapdh expression was affected by the TP53 mutation, which has been observed previously (Hölzl-Armstrong et al, 2020). β-Actin was included as a second loading control and its expression was constant for all samples, indicating accurate gel loading.…”

Section: Expression and Induction Of P53 Pathway Proteins In N-oh-4supporting

confidence: 59%

“…Cell viability was assessed by crystal violet staining as described previously (Hölzl‐Armstrong et al ., 2020) for concentrations up to 150 μM N ‐OH‐4‐ABP diluted in growth medium. In addition to a 24 hr time‐point a 24 hr + 24 hr time‐point was included in which the treatment medium was replaced by fresh growth medium after 24 hr.…”

Section: Methodsmentioning

confidence: 99%

“…To evaluate the expression of DDR proteins whole cell lysates were prepared and Western blotting performed as described previously (Hölzl‐Armstrong et al ., 2020). The following antibodies were utilized: anti‐phospho‐p53 (Ser15; 1:2,000; Cell Signalling, Danvers, MA, #9284;), anti‐p53 (1:500; Cell Signalling, 2524S), anti‐p21 (1:2,000; BD Biosciences, Franklin Lakes, NJ, #BD556431), anti‐γ‐H2ax (Ser139; 1:1,000; Cell Signalling #9718), anti‐phospho‐Chk1 (Ser345; 1:1,000; Cell Signaling #2348), glyceraldehyde 3‐phosphate dehydrogenase (Gapdh; 1:25,000; Chemicon International, Temecula, CA, #MAB374), anti‐Mdm2 (1:750; Abcam, Cambridge, United Kingdom #ab16895) and anti‐β‐actin (1:25,000; Abcam #ab6276).…”

Section: Methodsmentioning

confidence: 99%

“…To assess lacZ mutagenicity pellets of treated HUFs were prepared and the mutation assay performed as described previously (Hölzl‐Armstrong et al ., 2020). Briefly, primary HUFs were seeded into 75‐cm 2 flasks and treated for 24 hr.…”

Section: Methodsmentioning

confidence: 99%

“…Importantly, three mutations (C135, G245 and D281) have previously been predominantly observed in untreated spontaneously immortalized HUF clones, and it cannot be ruled out that these mutations are unspecific to N-OH-4-ABP treatment but rather happened due to spontaneous mutation events. In addition, even though the spontaneous TP53 mutation frequency is very low (0-3.7%) in primary HUFs prepared and treated at 3% oxygen (Kucab et al, 2015;Kucab et al, 2016;Hölzl-Armstrong et al, 2020), some mutations could be depending on the conformation of the DNA, mispair with either adenine or guanine to result in G>T/C>A and G>C/C>G transversions, respectively (Besaratinia and Tommasi, 2013). Therefore, the predominant mutation type detected in the present study agrees with dG-C8-4-ABP adduct formation as it was detected in N-OH- (Marnett, 2000).…”

Section: Mutations Induced By 4-abp and Its Metabolites In The Litementioning

confidence: 99%

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Mutagenicity of N‐hydroxy‐4‐aminobiphenyl in human TP53 knock‐in (Hupki) mouse embryo fibroblasts

Hölzl-Armstrong

Kucab

Zwart

et al. 2021

Environ and Mol Mutagen

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TP53 harbors somatic mutations in more than half of human tumors with some showing characteristic mutation spectra that have been linked to environmental exposures. In bladder cancer, a unique distribution of mutations amongst several codons of TP53 has been hypothesized to be caused by environmental carcinogens including 4-aminobiphenyl (4-ABP). 4-ABP undergoes metabolic activation to N-hydroxy-4-aminobiphenyl (N-OH-4-ABP) and forms pre-mutagenic adducts in DNA, of which N-(deoxyguanosin-8-yl)-4-ABP (dG-C8-4-ABP) is the major one. Human TP53 knockin mouse embryo fibroblasts (HUFs) are a useful model to study the influence of environmental carcinogens on TP53-mutagenesis. By performing the HUF immortalization assay (HIMA) TP53-mutant HUFs are generated and mutations can be identified by sequencing. Here we studied the induction of mutations in human TP53 after treatment of primary HUFs with N-OH-4-ABP. In addition, mutagenicity in the bacterial lacZ reporter gene and the formation of dG-C8-4-ABP, measured by 32 P-postlabelling analysis, were determined in N-OH-4-ABP-treated primary HUFs. A total of 6% TP53-mutants were identified after treatment with 40 μM N-OH-4-ABP for 24 hr (n = 150) with G>C/C>G transversion being the main mutation type. The mutation spectrum found in the TP53 gene of immortalized N-OH-4-ABP-treated HUFs was unlike the one found in human bladder cancer. DNA adduct formation (40 adducts/10 8 nucleotides) was detected after 24 hr treatment with 40 μM N-OH-4-ABP, but lacZ mutagenicity was not observed. Adduct levels decreased substantially (sixfold) after a 24 hr recovery period indicating that primary HUFs can efficiently repair the dG-C8-4-ABP adduct possibly before mutations are fixed. In conclusion, the observed difference in the N-OH-4-ABP-induced TP53 mutation spectrum to that observed in human bladder tumors do not support a role of 4-ABP in human bladder cancer development.

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Section: Expression and Induction Of P53 Pathway Proteins In N-oh-4supporting

confidence: 59%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Mutations Induced By 4-abp and Its Metabolites In The Litementioning

confidence: 99%

See 3 more Smart Citations

Mutagenicity of N‐hydroxy‐4‐aminobiphenyl in human TP53 knock‐in (Hupki) mouse embryo fibroblasts

Hölzl-Armstrong

Kucab

Zwart

et al. 2021

Environ and Mol Mutagen

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Evidence for etiologic field changes in tongue distant from tumor in patients with squamous cell carcinoma of the oral tongue

Wang

Coates

et al. 2022

The Journal of Pathology

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Oral cancer is a paradigm of Slaughter's concept of field cancerization, where tumors are thought to originate within an area of cells containing genetic alterations that predispose to cancer development. The field size is unclear but may represent a large area of tissue, and the origin of mutations is also unclear. Here, we analyzed whole exome and transcriptome features in contralateral tumor-distal tongue (i.e. distant from the tumor, not tumor-adjacent) and corresponding tumor tissues of 15 patients with squamous cell carcinoma of the oral tongue. The number of point mutations ranged from 41 to 237 in tumors and from one to 78 in tumor-distal samples. Tumor-distal samples showed mainly clock-like (associated with aging) or tobacco smoking mutational signatures. Tumors additionally showed mutations that associate with cytidine deaminase AID/APOBEC enzyme activities or a UV-like signature. Importantly, no point mutations were shared between a tumor and the matched tumor-distal sample in any patient. TP53 was the most frequently mutated gene in tumors (67%), whereas a TP53 mutation was detected in only one tumor-distal sample, and this mutation was not shared with the matched tumor. Arm-level copy number variation (CNV) was found in 12 tumors, with loss of chromosome (Chr) 8p or gain of 8q being the most frequent events. Two tumor-distal samples showed a gain of Chr8, which was associated with increased expression of Chr8-located genes in these samples, although gene ontology did not show a role for these genes in oncogenic processes. In situ hybridization revealed a mixed pattern of Chr8 gain and neutral copy number in both tumor cells and adjacent nontumor epithelium in one patient. We conclude that distant field cancerization exists but does not present as tumor-related mutational events. The data are compatible with etiologic field effects, rather than classical monoclonal field cancerization theory.

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Association between acrylamide exposure and sex hormones among premenopausal and postmenopausal women: NHANES, 2013–2016

Wang

Deng

et al. 2023

J Endocrinol Invest

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Mutagenicity of acrylamide and glycidamide in human TP53 knock-in (Hupki) mouse embryo fibroblasts

Cited by 26 publications

References 51 publications

Mutagenicity of N‐hydroxy‐4‐aminobiphenyl in human TP53 knock‐in (Hupki) mouse embryo fibroblasts

Mutagenicity of N‐hydroxy‐4‐aminobiphenyl in human TP53 knock‐in (Hupki) mouse embryo fibroblasts

Evidence for etiologic field changes in tongue distant from tumor in patients with squamous cell carcinoma of the oral tongue

Association between acrylamide exposure and sex hormones among premenopausal and postmenopausal women: NHANES, 2013–2016

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