https://scholars.lib.ntu.edu.tw/handle/123456789/469963
標題: | Calreticulin regulates MYCN expression to control neuronal differentiation and stemness of neuroblastoma | 作者: | Lee, A.C.-L. Shih, Y.-Y. Zhou, F. Chao, T.-C. Lee, H. Liao, Y.-F. WEN-MING HSU HSIN-YU LEE |
公開日期: | 2019 | 出版社: | Springer Verlag | 卷: | 97 | 期: | 3 | 起(迄)頁: | 325-339 | 來源出版物: | Journal of Molecular Medicine | 摘要: | Oncogenic N-MYC (MYCN) is widely used as a biomarker in clinics for neuroblastoma (NB) patients; nevertheless, mechanism that underlines MYCN regulation remains elusive. In the present study, we identified calreticulin (CRT) as a novel MYCN suppressor that downregulated MYCN promoter activity and protein expression to modulate neuronal differentiation and stemness. Our data showed that CRT-mediated MYCN suppression led to increased neurite length and commensurate elevation in differentiation marker GAP-43. We examined effect of radiotherapy and discovered that ionizing radiation (IR) was able to augment CRT expression dose-dependently in NB. Interestingly, neuronal differentiation and neurosphere formation (NSF) of NB were not only co-modulated by IR and CRT but were also dependent on Ca 2+ -buffering domain (C-domain) of CRT. Mutagenesis analysis showed that C-domain was indispensable for CRT-mediated MYCN regulation in NB differentiation and NSF. Of note, IR-induced formation of neural stem-like neurospheres (NS) was significantly impaired in CRT-overexpressed NB cells. The occupancy of CRT on MYCN 5′ proximal promoter was confirmed by chromatin immunoprecipitation assays, revealing potential CRT binding sites that coincided with transcription factor E2F1 binding elements. In addition, we identified a physical interaction between CRT and E2F1, and demonstrated that CRT occupancy on MYCN promoter prevented E2F1-mediated MYCN upregulation. In line with in vitro findings, hampered tumor latency and retarded tumor growth in xenograft model corroborated IR and CRT co-mediated neuronal differentiation of NB. Together, our data delineated a novel mechanism of CRT-mediated MYCN regulation and warranted further preclinical investigation towards new therapeutic strategy for NB. CRT suppresses MYCN expression and promotes neuronal differentiation in NB. CRT regulates MYCN via interaction with E2F1 and direct binding to MYCN promoter. Ca 2+ -buffering domain of CRT is critical in MYCN regulation and NB differentiation. CRT-MYCN axis impacts on NB stemness by modulating neurosphere formation. Xenograft model corroborates in vitro NB differentiation mediated by CRT and IR. ? 2019, Springer-Verlag GmbH Germany, part of Springer Nature. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85059591909&doi=10.1007%2fs00109-018-1730-x&partnerID=40&md5=cd04c3689a10fa8f89737a1e48a31b33 https://scholars.lib.ntu.edu.tw/handle/123456789/469963 |
ISSN: | 0946-2716 | DOI: | 10.1007/s00109-018-1730-x | SDG/關鍵字: | calcium ion; calreticulin; N Myc proto oncogene protein; neuromodulin; transcription factor E2F1; calreticulin; calreticulin, human; MYCN protein, human; N Myc proto oncogene protein; 5' untranslated region; animal experiment; animal model; Article; binding site; cancer patient; cancer radiotherapy; cancer stem cell; chromatin immunoprecipitation; clinical article; cohort analysis; controlled study; gene overexpression; human; human cell; human tissue; in vitro study; in vivo study; ionizing radiation; mouse; mutagenesis; nerve cell differentiation; neurite; neurite outgrowth; neuroblastoma; neuroblastoma cell line; nonhuman; oncogene c myb; promoter region; protein expression; protein protein interaction; receptor down regulation; SK-N-BE(2) cell line; SK-N-SH cell line; tumor growth; tumor suppressor gene; tumor xenograft; animal; female; genetics; metabolism; nerve cell; nervous system development; neuroblastoma; pathology; physiology; SCID mouse; tumor cell line; Animals; Calreticulin; Cell Line, Tumor; Female; Humans; Mice, SCID; N-Myc Proto-Oncogene Protein; Neuroblastoma; Neurogenesis; Neurons; Promoter Regions, Genetic; Radiation, Ionizing |
顯示於: | 醫學系 |
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