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TOP2B Is Required to Maintain the Adrenergic Neural Phenotype and for ATRA‑Induced Differentiation of SH‑SY5Y Neuroblastoma Cells

Lookup NU author(s): Mushtaq Khazeem, John CasementORCiD, Dr Niall Kenneth, Jade McGow, Dr Ian Cowell, Professor Caroline AustinORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

The neuroblastoma cell line SH-SY5Y is widely used to study retinoic acid (RA)-induced gene expression and differentiation and as a tool to study neurodegenerative disorders. SH-SY5Y cells predominantly exhibit adrenergic neuronal properties, but they can also exist in an epigenetically interconvertible alternative state with more mesenchymal characteristics; as a result, these cells can be used to study gene regulation circuitry controlling neuroblastoma phenotype. Using a combination of pharmacological inhibition and targeted gene inactivation, we have probed the requirement for DNA topoisomerase IIB (TOP2B) in RA-induced gene expression and differentiation and in the balance between adrenergic neuronal versus mesenchymal transcription programmes. We found that expression of many, but not all genes that are rapidly induced by ATRA in SH-SY5Y cells was significantly reduced in the TOP2B null cells; these genes include BCL2, CYP26A1, CRABP2, and NTRK2. Comparing gene expression profiles in wild-type versus TOP2B null cells, we found that long genes and genes expressed at a high level in WT SH-SY5Y cells were disproportionately dependent on TOP2B. Notably, TOP2B null SH-SY5Y cells upregulated mesenchymal markers vimentin (VIM) and fibronectin (FN1) and components of the NOTCH signalling pathway. Enrichment analysis and comparison with the transcription profiles of other neuroblastoma-derived cell lines supported the conclusion that TOP2B is required to fully maintain the adrenergic neural-like transcriptional signature of SH-SY5Y cells and to suppress the alternative mesenchymal epithelial-like epigenetic state.


Publication metadata

Author(s): Khazeem mm, Casement JW, Scholssmacher G, Kenneth NS, Sumbung NK, Chan JYT, McGow JF, Cowell IG, Austin CA

Publication type: Article

Publication status: Published

Journal: Molecular Neurobiology

Year: 2022

Volume: 59

Online publication date: 13/07/2022

Acceptance date: 23/06/2022

Date deposited: 25/07/2022

ISSN (print): 0893-7648

ISSN (electronic): 1559-1182

Publisher: Springer

URL: https://doi.org/10.1007/s12035-022-02949-6

DOI: 10.1007/s12035-022-02949-6

PubMed id: 35831557


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Funding

Funder referenceFunder name
the Higher Committee for Education Development in Iraq

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