| 研究実績の概要 |
ATRX is a tumor suppressor gene, and its recurrent somatic mutations relate to older neuroblastoma (NB) patients with advanced stages. Recent reports indicate that ATRX deficiency has been linked to replication stress and DNA damage by way of G-quadruplex (G4) DNA secondary structure. Here we found that NB cells lacking ATRX depends on p53 deficiency to limit replication–fork progression and genomic instability.
Previously, we knocked out (KO) ATRX in MYCN amplified (NGP) and MYCN single copy (SK-N-AS) NB cells with wild type and mutant TP53, respectively, using CRISPR-Cas9 system. By in vitro analysis, ATRX loss results in increased expression of gamma-H2AX, indicating the accumulation of endogenous DNA damage in the ATRX KO NGP cells but not in SK-N-AS clones. Using a monoclonal antibody known to recognize G4 structures (1H6), we found that ATRX loss increased G4 formation only in ATRX KO NGP cells. Furthermore, lentiviral-mediated inactivation of p53 reduced the G4 accumulation in the ATRX KO NGP cells, suggesting that ATRX plays a role in maintaining genome integrity and p53 deficiency helps to release replication stress and DNA damage in NB cells featuring inactivated ATRX.
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| 次年度使用額が生じた理由 |
Because of novel corona virus, we couldn't present our study in different meetings. And also some experiment materials, we couldn't buy last fiscal time.
This year, we are going to submit our manuscript and add the remaining
amount in new fiscal year for article cost. And try to attend the national conference to present our data.
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