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PAX3-NCOA2 fusion gene has a dual role in promoting the proliferation and inhibiting the myogenic differentiation of rhabdomyosarcoma cells

Oncogene volume 33, pages 5601–5608 (2014)Cite this article

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Abstract

We analyzed a complex chromosomal translocation in a case of embryonal rhabdomyosarcoma (RMS) and showed that it generates the fusion gene PAX3 (paired box 3)-NCOA2 (nuclear receptor coactivator 2). To understand the role of this translocation in RMS tumorigenesis, we established two types of stable mouse myoblast C2C12 cell lines expressing PAX3-NCOA2 and PAX3-FOXO1A (forkhead box O1A), respectively. Compared with control cells, PAX3-NCOA2 cells grew faster, were more motile, were less anchorage dependent, progressed more quickly through the G1/S phase of cell cycle and showed greater transcriptional activation of the PAX3 consensus-binding site. However, PAX3-NCOA2 cells proliferated more slowly and differentiated more weakly than did PAX3-FOXO1A cells. Both PAX3-NCOA2 cells and PAX3-FOXO1A cells formed tumors in nude mice, although the PAX3-NCOA2-induced tumors grew more slowly. Our results may explain why NCOA2 rearrangement is mainly found in embryonal rhabdomyosarcoma, which has a better prognosis than alveolar rhabdomyosarcoma, which expresses the PAX3-FOXO1A fusion gene. These results indicate that the PAX3-NCOA2 fusion gene has a dual role in the tumorigenesis of RMS: promotion of the proliferation and inhibition of the myogenic differentiation of RMS cells.

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Abbreviations

ARMS:

alveolar rhabdomyosarcoma

BAC:

bacterial artificial chromosome

bHLH:

sequence similarity with basic helix–loop–helix motifs

CBP:

CREB-binding protein

cDNA:

complementary DNA

CID:

CBP interaction domain

CREB:

cAMP (cyclic adenosine monophosphate) response element-binding protein

DM:

differentiation medium

ERMS:

embryonal rhabdomyosarcoma

FISH:

fluorescence in situ hybridization

FKHR:

forkhead in human rhabdomyosarcoma

FOXO1A:

forkhead box O1A

G1:

Gap1

GFP:

green fluorescent protein

H&E:

hematoxylin and eosin

LXD:

LXXLL-containing helical motif

MHC:

myosin heavy chain

MSCV:

murine stem cell virus

NCOA2:

nuclear receptor coactivator 2

NID:

nuclear receptor-interacting domain

PAS:

sequence similarity with the Per Arndt-Sim (PAS) motifs

PAX3:

paired box 3

PBS:

phosphate-buffered saline

PBST:

phosphate-buffered saline with Tween 20

PI:

propidium iodide

RMS:

rhabdomyosarcoma

RT–PCR:

reverse transcription–polymerase chain reaction

SDS–PAGE:

sodium dodecylsulfate–polyacrylamidegel electrophoresis

TAD:

transactivation domain

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Acknowledgements

This work was supported by Grant-in-aid for Scientific Research (A) 21249085, (A) 25253095, (B) 22390239, (C) 22591166, Grant-in-aid for Young Scientists (B) 24791078 and Grant-in-aid for Project for Development of Innovative Research on Cancer Therapeutics from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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  1. H Yoshida and M Miyachi: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pediatrics, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan

    H Yoshida, M Miyachi, K Sakamoto, K Ouchi, S Yagyu, K Kikuchi, Y Kuwahara, K Tsuchiya, T Imamura, T Iehara & H Hosoi

  2. Department of Environmental and Preventive Medicine, Shimane University School of Medicine, Shimane, Japan

    N Kakazu

  3. First Department of Pathology, Fukushima Medical University School of Medicine, Fukushima, Japan

    H Hojo

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Yoshida, H., Miyachi, M., Sakamoto, K. et al. PAX3-NCOA2 fusion gene has a dual role in promoting the proliferation and inhibiting the myogenic differentiation of rhabdomyosarcoma cells. Oncogene 33, 5601–5608 (2014). https://doi.org/10.1038/onc.2013.491

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