Abstract
Chromosome aberrations (CAs), i.e. changes in chromosome number or structure, are known to cause chromosome rearrangements and subsequently tumorigenesis. However, the involvement of CAs in chemical-induced carcinogenesis is unclear. In the current study, we aimed to clarify the possible involvement of CAs in chemical carcinogenesis using a rat model with the non-mutagenic hepatocarcinogen acetamide. In an in vivo micronucleus (MN) test, acetamide was revealed to induce CAs specifically in rat liver at carcinogenic doses. Acetamide also induced centromere-positive large MN (LMN) in hepatocytes. Immunohistochemical and electron microscopic analyses of the LMN, which can be histopathologically detected as basophilic cytoplasmic inclusion, revealed abnormal expression of nuclear envelope proteins, increased heterochromatinization, and massive DNA damage. These molecular pathological features in LMN progressed with acetamide exposure in a time-dependent manner, implying that LMN formation can lead to chromosome rearrangements. Overall, these data suggested that CAs induced by acetamide play a pivotal role in acetamide-induced hepatocarcinogenesis in rats and that CAs can cause chemical carcinogenesis in animals via MN formation.
Similar content being viewed by others
References
Abbott PJ, Mattia A, Renwick AJ, DiNovi M (2006) Aliphatic and aromatic amines and amides. Saf Eval Certain Food Addit 327–402
Bakhoum SF, Ngo B, Laughney AM et al (2018) Chromosomal instability drives metastasis through a cytosolic DNA response. Nature 553:467–472. https://doi.org/10.1038/nature25432
Cortés-Ciriano I, Lee JJK, Xi R et al (2020) Comprehensive analysis of chromothripsis in 2,658 human cancers using whole-genome sequencing. Nat Genet. https://doi.org/10.1038/s41588-019-0576-7
Crasta K, Ganem NJ, Dagher R et al (2012) DNA breaks and chromosome pulverization from errors in mitosis. Nature 482:53–58. https://doi.org/10.1038/nature10802
de Stoppelaar JM, de Roos B, Mohn GR, Hoebee B (1997) Analysis of DES-induced micronuclei in binucleated rat fibroblasts: comparison between FISH with a rat satellite I probe and immunocytochemical staining with CREST serum. Mutat Res 392:139–149. https://doi.org/10.1016/s0165-1218(97)00052-9
Dewhurst SM (2020) Chromothripsis and telomere crisis: engines of genome instability. Curr Opin Genet Dev 60:41–47. https://doi.org/10.1016/j.gde.2020.02.009
Fleischman RW, Baker JR, Hagopian M et al (1980) Carcinogenesis bioassay of acetamide, hexanamide, adipamide, urea and P-tolylurea in mice and rats. J Environ Pathol Toxicol 3:149–170
Guo X, Ni J, Liang Z et al (2019) The molecular origins and pathophysiological consequences of micronuclei: new insights into an age-old problem. Mutat Res 779:1–35. https://doi.org/10.1016/j.mrrev.2018.11.001
Halfmann CT, Sears RM, Katiyar A et al (2019) Repair of nuclear ruptures requires barrier-to-autointegration factor. J Cell Biol 218:2136–2149. https://doi.org/10.1083/jcb.201901116
Hamada S, Shigano M, Kawakami S et al (2019) Evaluation of the novel liver micronucleus assay using formalin-fixed tissues. Genes Environ 41:1–8. https://doi.org/10.1186/s41021-019-0128-5
Hatch EM (2018) Nuclear envelope rupture: little holes, big openings. Curr Opin Cell Biol 52:66–72. https://doi.org/10.1016/j.ceb.2018.02.001
Hatch EM, Hetzer MW (2015) Linking micronuclei to chromosome fragmentation. Cell 161:1502–1504. https://doi.org/10.1016/j.cell.2015.06.005
Hatch EM, Fischer AH, Deerinck TJ, Hetzer MW (2013) Catastrophic nuclear envelope collapse in cancer cell micronuclei. Cell 154:47. https://doi.org/10.1016/j.cell.2013.06.007
Jackson B, Dessau FI (1961) Liver tumors in rats fed acetamide. Lab Invest 10:909–923
Kastenbaum MA, Bowman KO (1970) Tables for determining the statistical significance of mutation frequencies. Mutat Res 9:527–549. https://doi.org/10.1016/0027-5107(70)90038-2
Kneissig M, Keuper K, De Pagter MS et al (2019) Micronuclei-based model system reveals functional consequences of chromothripsis in human cells. Elife 8:1–20. https://doi.org/10.7554/eLife.50292
Liu S, Kwon M, Mannino M et al (2018) Nuclear envelope assembly defects link mitotic errors to chromothripsis. Nature 561:551–555. https://doi.org/10.1038/s41586-018-0534-z
Maillet V, Boussetta N, Leclerc J et al (2018) LKB1 as a gatekeeper of hepatocyte proliferation and genomic integrity during liver regeneration. Cell Rep 22:1994–2005. https://doi.org/10.1016/j.celrep.2018.01.086
Misteli T, Soutoglou E (2009) The emerging role of nuclear architecture in DNA repair and genome maintenance. Nat Rev Mol Cell Biol 10:243–254. https://doi.org/10.1038/nrm2651
Moore MM, Gollapudi B, Nagane R et al (2019) The food contaminant acetamide is not an in vivo clastogen, aneugen, or mutagen in rodent hematopoietic tissue. Regul Toxicol Pharmacol 108:104451. https://doi.org/10.1016/j.yrtph.2019.104451
Nakamura K, Ishii Y, Takasu S et al (2020) Lack of in vivo mutagenicity of acetamide in a 13-week comprehensive toxicity study using F344 gpt delta rats. Toxicol Sci 177:431–440. https://doi.org/10.1093/toxsci/kfaa126
OECD (2016) OECD Guidline for the Testing of Chemicals. Test No. 489: In Vivo Mammalian Alkaline Comet Assay. https://read.oecd-ilibrary.org/environment/test-no-489-in-vivo-mammalian-alkaline-comet-assay_9789264264885-en
Pellestor F, Gatinois V (2020) Chromoanagenesis: a piece of the macroevolution scenario. Mol Cytogenet 13:1–9. https://doi.org/10.1186/s13039-020-0470-0
Qi R, Xu N, Wang G et al (2015) The lamin-A/C-LAP2α-BAF1 protein complex regulates mitotic spindle assembly and positioning. J Cell Sci 128:2830–2841. https://doi.org/10.1242/jcs.164566
Ranade D, Pradhan R, Jayakrishnan M et al (2019) Lamin A/C and Emerin depletion impacts chromatin organization and dynamics in the interphase nucleus. BMC Mol Cell Biol 20:1–20. https://doi.org/10.1186/s12860-019-0192-5
Russo A, Degrassi F (2018) Molecular cytogenetics of the micronucleus: still surprising. Mutat Res Genet Toxicol Environ Mutagen 836:36–40. https://doi.org/10.1016/j.mrgentox.2018.05.011
Santaguida S, Amon A (2015) Short- and long-term effects of chromosome mis-segregation and aneuploidy. Nat Rev Mol Cell Biol 16:473–485. https://doi.org/10.1038/nrm4025
Shigano M, Takashima R, Takasawa H, Hamada S (2016) Optimization of specimen preparation from formalin-fixed liver tissues for liver micronucleus assays: Hepatocyte staining with fluorescent dyes. Mutat Res Genet Toxicol Environ Mutagen 800–801:35–39. https://doi.org/10.1016/j.mrgentox.2016.03.004
Smith ER, Capochichi CD, Xu XX (2018) Defective nuclear lamina in aneuploidy and carcinogenesis. Front Oncol 8:1–10. https://doi.org/10.3389/fonc.2018.00529
Stephens PJ, Greenman CD, Fu B et al (2011) Massive genomic rearrangement acquired in a single catastrophic event during cancer development. Cell 144:27–40. https://doi.org/10.1016/j.cell.2010.11.055
Takeiri A, Motoyama S, Matsuzaki K et al (2013) New DNA probes to detect aneugenicity in rat bone marrow micronucleated cells by a pan-centromeric FISH analysis. Mutat Res Genet Toxicol Environ Mutagen 755:73–80. https://doi.org/10.1016/j.mrgentox.2013.05.011
Tang Z, Yang J, Wang X et al (2018) Active DNA end processing in micronuclei of ovarian cancer cells. BMC Cancer 18:1–10. https://doi.org/10.1186/s12885-018-4347-0
Terradas M, Martín M, Genescà A (2016) Impaired nuclear functions in micronuclei results in genome instability and chromothripsis. Arch Toxicol 90:2657–2667. https://doi.org/10.1007/s00204-016-1818-4
Tijhuis AE, Johnson SC, McClelland SE (2019) The emerging links between chromosomal instability (CIN), metastasis, inflammation and tumour immunity. Mol Cytogenet. https://doi.org/10.1186/s13039-019-0429-1
Tweats D, Eastmond DA, Lynch AM et al (2019) Role of aneuploidy in the carcinogenic process: Part 3 of the report of the 2017 IWGT workgroup on assessing the risk of aneugens for carcinogenesis and hereditary diseases. Mutat Res Genet Toxicol Environ Mutagen 847:403032. https://doi.org/10.1016/j.mrgentox.2019.03.005
Uno Y, Morita T, Luijten M et al (2015a) Micronucleus test in rodent tissues other than liver or erythrocytes: report of the IWGT working group. Mutat Res Genet Toxicol Environ Mutagen 783:19–22. https://doi.org/10.1016/j.mrgentox.2015.03.001
Uno Y, Morita T, Luijten M et al (2015b) Recommended protocols for the liver micronucleus test: report of the IWGT working group. Mutat Res Genet Toxicol Environ Mutagen 783:13–18. https://doi.org/10.1016/j.mrgentox.2014.10.010
van Steensel B, Belmont AS (2017) Lamina-associated domains: links with chromosome architecture, heterochromatin, and gene repression. Cell 169:780–791. https://doi.org/10.1016/j.cell.2017.04.022
Vismeh R, Haddad D, Moore J et al (2018) Exposure assessment of acetamide in milk, beef, and coffee using xanthydrol derivatization and gas chromatography/mass spectrometry. J Agric Food Chem 66:298–305. https://doi.org/10.1021/acs.jafc.7b02229
Voronina N, Wong JKL, Hübschmann D et al (2020) The landscape of chromothripsis across adult cancer types. Nat Commun 11:2320. https://doi.org/10.1038/s41467-020-16134-7
Zhang CZ, Spektor A, Cornils H et al (2015) Chromothripsis from DNA damage in micronuclei. Nature 522:179–184. https://doi.org/10.1038/nature14493
Zhang S, Zhou K, Luo X et al (2018) The polyploid state plays a tumor-suppressive role in the liver. Dev Cell 44:447-459.e5. https://doi.org/10.1016/j.devcel.2018.01.010
Acknowledgements
We thank Ms. Ayako Saikawa and Ms. Yoshimi Komatsu for expert technical assistance in processing histological materials.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
Kenji Nakamura is an employee of Ono Pharmaceutical Co., Ltd., Osaka, Japan. The authors declare no conflicts of interest associated with this manuscript.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Nakamura, K., Ishii, Y., Takasu, S. et al. Chromosome aberrations induced by the non-mutagenic carcinogen acetamide involve in rat hepatocarcinogenesis through micronucleus formation in hepatocytes. Arch Toxicol 95, 2851–2865 (2021). https://doi.org/10.1007/s00204-021-03099-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-021-03099-9