Open AccessDownregulated NORAD in neuroblastoma promotes cell proliferation via chromosomal instability and predicts poor prognosis
Author(s) -
Yizhi Yu,
Feng Chen,
Yichen Jin,
Yuxue Yang,
Shengcai Wang,
Jie Zhang,
Chenghao Chen,
Qi Zeng,
Wei Han,
Huanmin Wang,
Yongli Guo
Publication year - 2020
Publication title -
acta biochimica polonica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.452
H-Index - 78
eISSN - 1734-154X
pISSN - 0001-527X
DOI - 10.18388/abp.2020_5454
Subject(s) - gene knockdown , biology , cell cycle , cancer research , gene silencing , chromosome instability , cell growth , genome instability , dna damage , dna repair , neuroblastoma , parp1 , cell , microbiology and biotechnology , genetics , gene , cell culture , dna , chromosome , poly adp ribose polymerase , polymerase
Increasing evidence suggests that long non-coding RNAs (lncRNAs) are involved in neuroblastoma (NB) pathogenesis. The aim of this study was to elucidate the roles and underlying mechanism of non-coding RNA activated by DNA damage (NORAD) in childhood NB. Both public data and clinical specimens were used to determine NORAD expression. Colony formation, cell proliferation and wound healing assays were performed to evaluate NORAD effects on proliferation and migration of SH-SY5Y and SK-N-BE(2) cells. Flow cytometry was used to examine the cell cycle changes. The expression of genes and proteins involved in chromosomal instability was determined by qRT-PCR and western blotting, respectively. Our results showed that low NORAD expression correlated with advanced tumor stage, high risk and MYCN amplification in both public data and clinical samples. Kaplan–Meier analysis indicated that patients with low NORAD expression had poor survival outcomes. Functional research showed that NORAD knockdown promoted cell proliferation and migration, and arrested the cell cycle at the G2/M phase. Moreover, the expression of the DNA damage sensor, PARP1, increased after NORAD knockdown, indicating a potential contribution of NORAD to DNA damage repair. NORAD silencing also affected the expression of genes and proteins related to sister chromatid cohesion and segregation, which are involved in chromosomal instability and consequent aneuploidy. These results suggest that NORAD may serve as a tumor suppressor in NB pathogenesis and progression. Thus, NORAD is a potential therapeutic target and a promising prognostic marker for NB patients.