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The long non-coding RNA MYCNOS-01 regulates MYCN protein levels and affects growth of MYCN-amplified rhabdomyosarcoma and neuroblastoma cells

The long non-coding RNA MYCNOS-01 regulates MYCN protein levels and affects growth of MYCN-amplified rhabdomyosarcoma and neuroblastoma cells
The long non-coding RNA MYCNOS-01 regulates MYCN protein levels and affects growth of MYCN-amplified rhabdomyosarcoma and neuroblastoma cells

BACKGROUND: MYCN is amplified in small cell lung cancers and several pediatric tumors, including alveolar rhabdomyosarcomas and neuroblastomas. MYCN protein is known to play a key oncogenic role in both alveolar rhabdomyosarcomas and neuroblastomas. MYCN opposite strand (MYCNOS) is a gene located on the antisense strand to MYCN that encodes alternatively spliced transcripts, two of which (MYCNOS-01 and MYCNOS-02) are known to be expressed in neuroblastoma and small cell lung cancer with reciprocal regulation between MYCNOS-02 and MYCN reported for neuroblastomas. We sought to determine a functional role for MYCNOS-01 in alveolar rhabdomyosarcoma and neuroblastoma cells and identify any associated regulatory effects between MYCN and MYCNOS-01.

METHODS: MYCNOS-01, MYCNOS-02 and MYCN expression levels were assessed in alveolar rhabdomyosarcoma and neuroblastoma cell lines and tumor samples from patients using Affymetrix microarray data and quantitative RT-PCR. Following MYCNOS-01 or MYCN siRNA knockdown and MYCNOS-01 overexpression, transcript levels were assayed by quantitative RT-PCR and MYCN protein expression assessed by Western blot and immunofluorescence. Additionally, effects on cell growth, apoptosis and cell cycle profiles were determined by a metabolic assay, caspase activity and flow cytometry, respectively.

RESULTS: MYCNOS-01 transcript levels were generally higher in NB and RMS tumor samples and cell lines with MYCN genomic amplification. RNA interference of MYCNOS-01 expression did not alter MYCN transcript levels but decreased MYCN protein levels. Conversely, MYCN reduction increased MYCNOS-01 transcript levels, creating a negative feedback loop on MYCN protein levels. Reduction of MYCNOS-01 or MYCN expression decreased cell growth in MYCN-amplified alveolar rhabdomyosarcoma and neuroblastoma cell lines. This is consistent with MYCNOS-01-mediated regulation of MYCN contributing to the phenotype observed.

CONCLUSIONS: An alternative transcript of MYCNOS, MYCNOS-01, post-transcriptionally regulates MYCN levels and affects growth in MYCN-amplified rhabdomyosarcoma and neuroblastoma cells.

Journal Article, Research Support, Non-U.S. Gov't
1471-2407
O'Brien, Eleanor M.
1e74780c-dc65-4d54-b651-bb066f906b8c
Selfe, Joanna L.
4e764d15-2225-45b5-9684-ea9f1ed2ddf3
Martins, Ana Sofia
9b7ee34c-76af-4f7a-9fd0-356544f23544
Walters, Zoë S.
e1ccd35d-63a9-4951-a5da-59122193740d
Shipley, Janet M.
24c1f3c9-c8d9-49d7-ab51-3ddd41abf975
O'Brien, Eleanor M.
1e74780c-dc65-4d54-b651-bb066f906b8c
Selfe, Joanna L.
4e764d15-2225-45b5-9684-ea9f1ed2ddf3
Martins, Ana Sofia
9b7ee34c-76af-4f7a-9fd0-356544f23544
Walters, Zoë S.
e1ccd35d-63a9-4951-a5da-59122193740d
Shipley, Janet M.
24c1f3c9-c8d9-49d7-ab51-3ddd41abf975

O'Brien, Eleanor M., Selfe, Joanna L., Martins, Ana Sofia, Walters, Zoë S. and Shipley, Janet M. (2018) The long non-coding RNA MYCNOS-01 regulates MYCN protein levels and affects growth of MYCN-amplified rhabdomyosarcoma and neuroblastoma cells. BMC cancer, 18 (1), [217]. (doi:10.1186/s12885-018-4129-8).

Record type: Article

Abstract

BACKGROUND: MYCN is amplified in small cell lung cancers and several pediatric tumors, including alveolar rhabdomyosarcomas and neuroblastomas. MYCN protein is known to play a key oncogenic role in both alveolar rhabdomyosarcomas and neuroblastomas. MYCN opposite strand (MYCNOS) is a gene located on the antisense strand to MYCN that encodes alternatively spliced transcripts, two of which (MYCNOS-01 and MYCNOS-02) are known to be expressed in neuroblastoma and small cell lung cancer with reciprocal regulation between MYCNOS-02 and MYCN reported for neuroblastomas. We sought to determine a functional role for MYCNOS-01 in alveolar rhabdomyosarcoma and neuroblastoma cells and identify any associated regulatory effects between MYCN and MYCNOS-01.

METHODS: MYCNOS-01, MYCNOS-02 and MYCN expression levels were assessed in alveolar rhabdomyosarcoma and neuroblastoma cell lines and tumor samples from patients using Affymetrix microarray data and quantitative RT-PCR. Following MYCNOS-01 or MYCN siRNA knockdown and MYCNOS-01 overexpression, transcript levels were assayed by quantitative RT-PCR and MYCN protein expression assessed by Western blot and immunofluorescence. Additionally, effects on cell growth, apoptosis and cell cycle profiles were determined by a metabolic assay, caspase activity and flow cytometry, respectively.

RESULTS: MYCNOS-01 transcript levels were generally higher in NB and RMS tumor samples and cell lines with MYCN genomic amplification. RNA interference of MYCNOS-01 expression did not alter MYCN transcript levels but decreased MYCN protein levels. Conversely, MYCN reduction increased MYCNOS-01 transcript levels, creating a negative feedback loop on MYCN protein levels. Reduction of MYCNOS-01 or MYCN expression decreased cell growth in MYCN-amplified alveolar rhabdomyosarcoma and neuroblastoma cell lines. This is consistent with MYCNOS-01-mediated regulation of MYCN contributing to the phenotype observed.

CONCLUSIONS: An alternative transcript of MYCNOS, MYCNOS-01, post-transcriptionally regulates MYCN levels and affects growth in MYCN-amplified rhabdomyosarcoma and neuroblastoma cells.

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s12885-018-4129-8 - Version of Record
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Accepted/In Press date: 13 February 2018
e-pub ahead of print date: 21 February 2018
Published date: 21 February 2018
Keywords: Journal Article, Research Support, Non-U.S. Gov't

Identifiers

Local EPrints ID: 421215
URI: http://eprints.soton.ac.uk/id/eprint/421215
ISSN: 1471-2407
PURE UUID: c449a217-f0a6-4923-9861-5dd0c39dedec
ORCID for Zoë S. Walters: ORCID iD orcid.org/0000-0002-1835-5868

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Date deposited: 24 May 2018 16:31
Last modified: 26 Nov 2021 03:12

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Contributors

Author: Eleanor M. O'Brien
Author: Joanna L. Selfe
Author: Ana Sofia Martins
Author: Zoë S. Walters ORCID iD
Author: Janet M. Shipley

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