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ZAR1 knockdown promotes the differentiation of human neuroblastoma cells by suppression of MYCN expression

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Abstract

Although DNA hypermethylation at non-promoter region of the Zygote arrest 1 (ZAR1) gene has been observed in many types of tumor, including neuroblastoma (NB), the role of this gene in tumor development and/or progression is unclear. One reason is that knowledge about the function of ZAR1 protein is limited. Although it has been reported that ZAR1 plays a crucial role in early embryogenesis and may act as a transcriptional repressor for some transcripts, the detailed mechanism is still elusive. In the present study, we analyzed public data of NB patients and found that higher expression levels of ZAR1 were significantly associated with a shorter survival period. Consistent with this result, ZAR1-depleted NB cells showed well-differentiated phenotypes with elongated neurites and upregulated expression of TRKA and RET, which are markers for differentiated NB. Moreover, the expression level of MYCN protein was markedly suppressed in ZAR1-depleted NB cells. MYCN-depleted cells showed similar phenotypes to ZAR1-depleted cells. The present findings indicate that ZAR1 has oncogenic effects in NB by suppressing cell differentiation via regulation of MYCN expression.

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References

  1. Maris JM, Hogarty MD, Bagatell R, Cohn SL. Neuroblastoma. Lancet. 2007;369(9579):2106–20.

    Article  CAS  PubMed  Google Scholar 

  2. Schwab M, Alitalo K, Klempnauer KH, Varmus HE, Bishop JM, Gilbert F, Brodeur G, Goldstein M, Trent J. Amplified DNA with limited homology to myc cellular oncogene is shared by human neuroblastoma cell lines and a neuroblastoma tumour. Nature. 1983;305(5931):245–8.

    Article  CAS  PubMed  Google Scholar 

  3. Dang CV. MYC on the path to cancer. Cell. 2012;149(1):22–35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Caron H. Allelic loss of chromosome 1 and additional chromosome 17 material are both unfavourable prognostic markers in neuroblastoma. Med Pediatr Oncol. 1995;24(4):215–21.

    Article  CAS  PubMed  Google Scholar 

  5. Lampert F, Rudolph B, Christiansen H, Franke F. Identical chromosome 1p breakpoint abnormality in both the tumor and the constitutional karyotype of a patient with neuroblastoma. Cancer Genet Cytogenet. 1988;34(2):235–9.

    Article  CAS  PubMed  Google Scholar 

  6. Shinojima Y, Terui T, Hara H, Kimura M, Igarashi J, Wang X, Kawashima H, Kobayashi Y, Muroi S, Hayakawa S, Esumi M, Fujiwara K, Ghosh S, Yamamoto T, Held W, Nagase H. Identification and analysis of an early diagnostic marker for malignant melanoma: ZAR1 intra-genic differential methylation. J Dermatol Sci. 2010;59(2):98–106.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Watanabe T, Yachi K, Ohta T, Fukushima T, Yoshino A, Katayama Y, Shinojima Y, Terui T, Nagase H. Aberrant hypermethylation of non-promoter zygote arrest 1 (ZAR1) in human brain tumors. Neurol Med Chir (Tokyo). 2010;50(12):1062–9.

    Article  Google Scholar 

  8. Takagi K, Fujiwara K, Takayama T, Mamiya T, Soma M, Nagase H. DNA hypermethylation of zygote arrest 1 (ZAR1) in hepatitis C virus positive related hepatocellular carcinoma. Springerplus. 2013;2(1):150.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Sugito K, Kawashima H, Yoshizawa S, Uekusa S, Hoshi R, Furuya T, Kaneda H, Hosoda T, Konuma N, Masuko T, Ohashi K, Ikeda T, Koshinaga T, Tomita R, Shinojima Y, Fujiwara K, Watanabe T, Held WA, Nagase H. Non-promoter DNA hypermethylation of zygote arrest 1 (ZAR1) in neuroblastomas. J Pediatr Surg. 2013;48(4):782–8.

    Article  PubMed  Google Scholar 

  10. Wu X, Viveiros MM, Eppig JJ, Bai Y, Fitzpatrick SL, Matzuk MM. Zygote arrest 1 (Zar1) is a novel maternal-effect gene critical for the oocyte-to-embryo transition. Nat Genet. 2003;33(2):187–91.

    Article  CAS  PubMed  Google Scholar 

  11. Wu X, Wang P, Brown CA, Zilinski CA, Matzuk MM. Zygote arrest 1 (Zar1) is an evolutionarily conserved gene expressed in vertebrate ovaries. Biol Reprod. 2003;69(3):861–7.

    Article  CAS  PubMed  Google Scholar 

  12. Yamamoto TM, Cook JM, Kotter CV, Khat T, Silva KD, Ferreyros M, Holt JW, Knight JD, Charlesworth A. Zar1 represses translation in Xenopus oocytes and binds to the TCS in maternal mRNAs with different characteristics than Zar2. Biochim Biophys Acta. 2013;1829(10):1034–46.

    Article  CAS  PubMed  Google Scholar 

  13. Brodeur GM. Neuroblastoma: biological insights into a clinical enigma. Nat Rev Cancer. 2003;3(3):203–16.

    Article  CAS  PubMed  Google Scholar 

  14. Peterson S, Bogenmann E. The RET and TRKA pathways collaborate to regulate neuroblastoma differentiation. Oncogene. 2004;23(1):213–25.

    Article  CAS  PubMed  Google Scholar 

  15. Thelie A, Papillier P, Pennetier S, Perreau C, Traverso JM, Uzbekova S, Mermillod P, Joly C, Humblot P, Dalbies-Tran R. Differential regulation of abundance and deadenylation of maternal transcripts during bovine oocyte maturation in vitro and in vivo. BMC Dev Biol. 2007;7:125.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Charlesworth A, Yamamoto TM, Cook JM, Silva KD, Kotter CV, Carter GS, Holt JW, Lavender HF, MacNicol AM, Ying WY, Wilczynska A. Xenopus laevis zygote arrest 2 (zar2) encodes a zinc finger RNA-binding protein that binds to the translational control sequence in the maternal Wee1 mRNA and regulates translation. Dev Biol. 2012;369(2):177–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Hu J, Wang F, Zhu X, Yuan Y, Ding M, Gao S. Mouse ZAR1-like (XM_359149) colocalizes with mRNA processing components and its dominant-negative mutant caused two-cell-stage embryonic arrest. Dev Dyn. 2010;239(2):407–24.

    Article  CAS  PubMed  Google Scholar 

  18. Seeger RC, Brodeur GM, Sather H, Dalton A, Siegel SE, Wong KY, Hammond D. Association of multiple copies of the N-myc oncogene with rapid progression of neuroblastomas. N Engl J Med. 1985;313(18):1111–6.

    Article  CAS  PubMed  Google Scholar 

  19. Look AT, Hayes FA, Shuster JJ, Douglass EC, Castleberry RP, Bowman LC, Smith EI, Brodeur GM. Clinical relevance of tumor cell ploidy and N-myc gene amplification in childhood neuroblastoma: a Pediatric Oncology Group study. J Clin Oncol. 1991;9(4):581–91.

    Article  CAS  PubMed  Google Scholar 

  20. Cascon A, Robledo M. MAX and MYC: a heritable breakup. Cancer Res. 2012;72(13):3119–24.

    Article  CAS  PubMed  Google Scholar 

  21. Henriksen JR, Haug BH, Buechner J, Tomte E, Lokke C, Flaegstad T, Einvik C. Conditional expression of retrovirally delivered anti-MYCN shRNA as an in vitro model system to study neuronal differentiation in MYCN-amplified neuroblastoma. BMC Dev Biol. 2011;11:1.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Chesler L, Schlieve C, Goldenberg DD, Kenney A, Kim G, McMillan A, Matthay KK, Rowitch D, Weiss WA. Inhibition of phosphatidylinositol 3-kinase destabilizes Mycn protein and blocks malignant progression in neuroblastoma. Cancer Res. 2006;66(16):8139–46.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Otto T, Horn S, Brockmann M, Eilers U, Schuttrumpf L, Popov N, Kenney AM, Schulte JH, Beijersbergen R, Christiansen H, Berwanger B, Eilers M. Stabilization of N-Myc is a critical function of Aurora A in human neuroblastoma. Cancer Cell. 2009;15(1):67–78.

    Article  CAS  PubMed  Google Scholar 

  24. Miao L, Yuan Y, Cheng F, Fang J, Zhou F, Ma W, Jiang Y, Huang X, Wang Y, Shan L, Chen D, Zhang J. Translation repression by maternal RNA binding protein Zar1 is essential for early oogenesis in zebrafish. Development. 2017;144(1):128–38.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Ms. A. Oguni for her excellent technical support and Ms. K. Tagata for her secretarial assistance. The present study was supported in part by JSPS KAKENHI Grant Number 20793772 to R.H.; MEXT-Supported Program for the Strategic Research Foundation at Private Universities (2011–2015) to N.F., H.N., M.S., T.K., and K.F.

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Authors and Affiliations

  1. Department of Pediatric Surgery, Nihon University School of Medicine, 30-1 Oyaguchi-Kamicho, Itabashi, Tokyo, 173-0032, Japan

    Yosuke Watanabe, Yoshiaki Ishizuka, Takayuki Hirano, Eri Nagasaki-Maeoka, Reina Hoshi, Shinsuke Yoshizawa, Shota Uekusa, Hiroyuki Kawashima, Kiminobu Sugito & Tsugumichi Koshinaga

  2. Department of Molecular Oncology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan

    Kenichi Shinohara

  3. Division of Nephrology, Hypertension and Endocrinology, Department of Medicine, Nihon University School of Medicine, Itabashi, Tokyo, 173-0032, Japan

    Noboru Fukuda

  4. Laboratory of Cancer Genetics, Chiba Cancer Center Research Institute, Chiba, 260-8717, Japan

    Hiroki Nagase

  5. Division of General Medicine, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-Kamicho, Itabashi, Tokyo, 173-8610, Japan

    Masayoshi Soma & Kyoko Fujiwara

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  1. Yosuke Watanabe
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  2. Yoshiaki Ishizuka
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  8. Hiroyuki Kawashima
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  11. Noboru Fukuda
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  13. Masayoshi Soma
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Correspondence to Tsugumichi Koshinaga or Kyoko Fujiwara.

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Watanabe, Y., Ishizuka, Y., Hirano, T. et al. ZAR1 knockdown promotes the differentiation of human neuroblastoma cells by suppression of MYCN expression. Med Oncol 34, 158 (2017). https://doi.org/10.1007/s12032-017-0999-x

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  • DOI: https://doi.org/10.1007/s12032-017-0999-x

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