The Role of MYCN in the Failure of MYCN Amplified Neuroblastoma Cell Lines to G1 Arrest After DNA Damage

20003 Background: Signaling through growth factor receptors is important in neuroblastoma pathogenesis. Chromosome 1p36 is commonly deleted in neuroblastoma tumors and is associated with a poor prognosis. UBE4B, a gene in 1p36, has been reported mutated in high- risk neuroblastoma. We have found a direct interaction between UBE4B and hrs, a protein required for epidermal growth factor receptor (EGFR) trafficking, suggesting a link between EGFR trafficking and neuroblastoma pathogenesis. We have analyzed the role of UBE4B in the EGFR pathway in neuroblastoma cell lines. Methods: The expression of UBE4B, hrs and EGFR were analyzed by quantitative Western blot in a panel of 7 human neuroblastoma cell lines (SHEP, SKNAS, SKNSH, KCNR, SY5Y, LA155N, NGP). EGFR degradation rates were determined by examining the kinetics of cellular EGFR depletion following a pulse of ligand. Results: UBE4B levels were lowest in SKNAS and highest in NGP cells. Hrs levels were lowest in SKNSH cells and higher in other cell lines. EGFR levels were lowest in NGP and KCNR and highest in SKNAS cells. UBE4B levels were correlated with known 1p deletions. EGFR degradation rates were slowest in SKNAS cells and therefore correlated with cellular UBE4B levels. The low degradation rates were correlated with high cellular levels of EGFR. Conclusions: Expression levels of UBE4B are correlated in neuroblastoma cell lines with chromosome 1p deletions. Cell lines with lower levels of UBE4B degrade EGFR at a markedly slower rate, correlated with higher cellular EGFR levels. We hypothesize that UBE4B affects cell growth by interacting with hrs, directing EGFR for degradation. In its absence the ability of a cell to sort growth factor receptors for degradation is inhibited, resulting in growth factor receptor overabundance and uncontrolled cell growth. These results support the testing of EGFR inhibitors in a future phase I trial for children with neuroblastoma. No significant financial relationships to disclose.

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The HDAC6/8/10 inhibitor TH34 induces DNA damage-mediated cell death in human high-grade neuroblastoma cell lines

Archives of Toxicology ◽

10.1007/s00204-018-2234-8 ◽

2018 ◽

Vol 92 (8) ◽

pp. 2649-2664 ◽

Cited By ~ 8

Author(s):

Fiona R. Kolbinger ◽

Emily Koeneke ◽

Johannes Ridinger ◽

Tino Heimburg ◽

Michael Müller ◽

...

Keyword(s):

Dna Damage ◽

Cell Death ◽

Cell Lines ◽

Neuroblastoma Cell ◽

High Grade ◽

Neuroblastoma Cell Lines

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NTRK1/TrkA Activation Overrides the G2/M-Checkpoint upon Irradiation

Cancers ◽

10.3390/cancers13236023 ◽

2021 ◽

Vol 13 (23) ◽

pp. 6023

Author(s):

Christina Hassiepen ◽

Aashish Soni ◽

Ines Rudolf ◽

Vivian Boron ◽

Sebastian Oeck ◽

...

Keyword(s):

Dna Damage ◽

Neuroblastoma Cell ◽

Human Neuroblastoma Cell ◽

Human Neuroblastoma ◽

Checkpoint Activation ◽

Neuronal Precursor Cells ◽

Damage Response ◽

High Doses ◽

Neuroblastoma Cell Lines

High expression of the receptor tyrosine kinase TrkA/NTRK1 is associated with a favorable outcome in several solid tumors of childhood including neuroblastoma. During development, TrkA/NTRK1 governs migration and differentiation of neuronal precursor cells, while it is associated with mitotic dysfunction and altered DNA damage response, among others, in neuroblastoma. Here, we used human neuroblastoma cell lines with inducible TrkA/NTRK1 expression to mechanistically explore the role of TrkA/NTRK1 signaling in checkpoint activation after DNA damage induced by ionizing radiation (IR). TrkA/NTRK1 activated cells showed increased short-term cell viability upon IR compared to vector control cells. This was accompanied by a deficient G2/M-checkpoint at both low (1 Gy) and high doses (4 Gy) of IR. In a tightly controlled setting, we confirmed that this effect was strictly dependent on activation of TrkA/NTRK1 by its ligand, nerve growth factor (NGF). TrkA/NTRK1-expressing cells displayed impaired ATM and CHK1 phosphorylation, resulting in stabilization of CDC25B. In line with these findings, ATM or ATR inhibition recapitulated the effects of TrkA/NTRK1 activation on the IR-induced G2/M-checkpoint. In conclusion, we here provide first evidence for a previously unrecognized function of NTRK signaling in checkpoint regulation and the response to IR.

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No evidence of DNA damage by co-exposure to extremely low frequency magnetic fields and aluminum on neuroblastoma cell lines

Mutation Research/Genetic Toxicology and Environmental Mutagenesis ◽

10.1016/j.mrgentox.2017.09.001 ◽

2017 ◽

Vol 823 ◽

pp. 11-21 ◽

Cited By ~ 6

Author(s):

Milena Villarini ◽

Angela Gambelunghe ◽

Daniela Giustarini ◽

Maria Vittoria Ambrosini ◽

Cristina Fatigoni ◽

...

Keyword(s):

Dna Damage ◽

Magnetic Fields ◽

Cell Lines ◽

Neuroblastoma Cell ◽

Low Frequency ◽

Extremely Low Frequency ◽

Neuroblastoma Cell Lines

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ROLE OF CASPASE 8 AS A DETERMINANT IN TRAIL SENSITIVITY OF NEUROBLASTOMA CELL LINES

Pediatric Hematology and Oncology ◽

10.3109/08880010903271663 ◽

2009 ◽

Vol 26 (8) ◽

pp. 549-559 ◽

Cited By ~ 2

Author(s):

Haixia Tong ◽

Chunwei Lu ◽

Yanmin Yang ◽

Jihong Zhang ◽

Jinhua Zhang

Keyword(s):

Cell Lines ◽

Neuroblastoma Cell ◽

Caspase 8 ◽

Neuroblastoma Cell Lines ◽

Trail Sensitivity

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Flow cytometry analysis of single-strand DNA damage in neuroblastoma cell lines using the F7-26 monoclonal antibody

Cytometry Part A ◽

10.1002/cyto.a.20458 ◽

2007 ◽

Vol 71A (11) ◽

pp. 951-960 ◽

Cited By ~ 8

Author(s):

Rita S. Grigoryan ◽

Bo Yang ◽

Nino Keshelava ◽

Jerry R. Barnhart ◽

C. Patrick Reynolds

Keyword(s):

Monoclonal Antibody ◽

Flow Cytometry ◽

Dna Damage ◽

Cell Lines ◽

Neuroblastoma Cell ◽

Single Strand ◽

Flow Cytometry Analysis ◽

Neuroblastoma Cell Lines

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Micronuclei and apoptosis in glioma and neuroblastoma cell lines and role of other lesions in the reconstruction of cellular radiosensitivity

Radiation and Environmental Biophysics ◽

10.1007/s00411-001-0121-8 ◽

2001 ◽

Vol 40 (4) ◽

pp. 295-300 ◽

Cited By ~ 10

Author(s):

J. M. Akudugu ◽

L. Böhm

Keyword(s):

Cell Lines ◽

Neuroblastoma Cell ◽

Cellular Radiosensitivity ◽

Neuroblastoma Cell Lines

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Role of ALK activation in the development and maintenance of the neoplastic phenotype in neuroblastoma

Journal of Clinical Oncology ◽

10.1200/jco.2009.27.15_suppl.10008 ◽

2009 ◽

Vol 27 (15_suppl) ◽

pp. 10008-10008

Author(s):

M. Regairaz ◽

F. Munier ◽

H. Sartelet ◽

V. Marty ◽

M. Castaing ◽

...

Keyword(s):

Cell Proliferation ◽

Antitumor Activity ◽

Cell Lines ◽

Neuroblastoma Cell ◽

Wild Type ◽

Primary Tumors ◽

Normal Tissues ◽

Anaplastic Lymphoma ◽

Neuroblastoma Cell Lines

10008 Background: Activating mutations of the Anaplastic Lymphoma Kinase (ALK) receptor could be responsible for most familial neuroblastoma cases and for up to 15% of somatic cases. The objective of the present study was to further investigate the role of ALK activation in neuroblastoma. Methods: Tissue microarrays were constructed containing 101 primary tumors and 56 paired normal tissues. Sections were immunostained with anti-ALK or anti-P-ALK antibodies, and with antibodies directed against the ALK ligands: PTN (Pleiotrophin) or MDK (Midkine). The Wilcoxon signed rank test was applied for comparison of paired data. Associations with prognostic factors were analyzed using t-tests. Effects of the ALK inhibitor TAE684 (Novartis) on cell proliferation and signaling was evaluated in wild-type or mutated ALK neuroblastoma cell lines and xenografts. Results: ALK was expressed in about 100% of tumors and normal tissues, while phospho-ALK was detected in 5% of normal tissues and 50% of tumors. Sequencing of the kinase domain of ALK showed that its phosphorylation was largely independent of mutations and we found that MDK and PTN ligands were expressed in 66% and 50% of tumors, respectively. Interestingly, ALK, P-ALK, and MDK were expressed at higher levels in tumors as compared with paired normal tissues (p < 0.0001), while PTN showed an inverse tendency, being more expressed in normal tissues (p = 0.07). In tumors, P-ALK was associated with good-prognosis factors, including favorable stages (p = 0.01), absence of MYCN amplification (p = 0.05) and a younger age at diagnosis (p = 0.03). Inhibition of cell proliferation by TAE684 was detectible in all neuroblastoma cell lines, regardless of ALK status. However, TAE684 failed to demonstrate antitumor activity in advanced stage neuroblastoma xenografts expressing either a wild-type or a mutated ALK. Interestingly, ALK pathway activation (P-STAT3, P-AKT) was weak or barely detectible in these xenografts. Conclusions: ALK activation occurs during neuroblastoma oncogenesis, along with a concomitant switch between the expressions of PTN and MDK. However, ALK may not be a relevant therapeutic target since in vivo inhibition showed no antitumor activity. [Table: see text]

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