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Characterisation of the p53 pathway in cell lines established from TH-MYCN transgenic mouse tumours

Lookup NU author(s): Dr Lindi Chen, Arman Esfandiari, Professor John Lunec, Professor Deborah Tweddle

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This is the final published version of an article that has been published in its final definitive form by Spandidos Publications, 2018.

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Abstract

Cell lines established from the TH-MYCN transgenic murine model of neuroblastoma are a valuable preclinical, immunocompetent, syngeneic model of neuroblastoma, for which knowledge of their p53 pathway status is important. Trp53 status and functional response to Nutlin-3 and ionising radiation (IR) were determined in 6 adherent TH-MYCN transgenic cell lines using Sanger sequencing, immunoblotting and flow cytometry. Sensitivity to structurally diverse MDM2 inhibitors (Nutlin-3, MI-63, RG7388 and NDD0005) were determined using XTT proliferations assays. 2/6 cell lines were Trp53 homozygous mutant (NHO2A and 844MYCN+/+) and 1/6 (282MYCN+/-) was Trp53 heterozygous mutant. For 1/6 cell lines (NHO2A), DNA from the corresponding primary tumour was found to be Trp53 wt. In all cases, the presence of a mutation was consistent with aberrant p53 signalling in response to Nutlin-3 and IR. In comparison to TP53 wt human neuroblastoma cells, Trp53 wt murine control and TH-MYCN cell lines were significantly less sensitive to growth inhibition mediated by MI-63 and RG7388. These murine Trp53 wt and mutant TH-MYCN cell lines are useful syngeneic immunocompetent neuroblastoma models, the former to test p53-dependent therapies in combination with immunotherapies, such as anti-GD2, and the latter as models of chemoresistant relapsed neuroblastoma when aberrations in the p53 pathway are more common. The spontaneous development of Trp53 mutations in 3 cell lines from TH-MYCN mice may have arisen from MYCN oncogenic driven and/or ex-vivo selection. The identified species-dependent selectivity of MI-63 and RG7388 should be considered when interpreting in vivo toxicity studies of MDM2 inhibitors.


Publication metadata

Author(s): Chen L, Esfandiari A, Reaves W, Vu A, Hogarty MD, Lunec J, Tweddle DA

Publication type: Article

Publication status: Published

Journal: International Journal of Oncology

Year: 2018

Volume: 52

Issue: 3

Pages: 967-977

Print publication date: 01/03/2018

Online publication date: 31/01/2018

Acceptance date: 12/12/2017

Date deposited: 08/02/2018

ISSN (print): 1019-6439

ISSN (electronic): 1791-2423

Publisher: Spandidos Publications

URL: https://doi.org/10.3892/ijo.2018.4261

DOI: 10.3892/ijo.2018.4261


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