Abstract
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.
MeSH terms
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Aurora Kinase B / genetics
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Aurora Kinase B / metabolism
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CDC2 Protein Kinase / genetics
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CDC2 Protein Kinase / metabolism
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Cell Cycle Proteins / genetics
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Cell Cycle Proteins / metabolism
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Cell Line, Tumor
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Cell Movement
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Cell Proliferation
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Centromere Protein A / genetics
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Centromere Protein A / metabolism
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Child
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Chondroitin Sulfate Proteoglycans / genetics
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Chondroitin Sulfate Proteoglycans / metabolism
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Chromosomal Instability*
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Chromosomal Proteins, Non-Histone / genetics
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Chromosomal Proteins, Non-Histone / metabolism
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Datasets as Topic
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Gene Expression Regulation, Neoplastic
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Humans
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N-Myc Proto-Oncogene Protein / genetics*
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N-Myc Proto-Oncogene Protein / metabolism
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Neoplasm Staging
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Nervous System Neoplasms / diagnosis
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Nervous System Neoplasms / genetics*
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Nervous System Neoplasms / mortality
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Nervous System Neoplasms / pathology
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Neuroblastoma / diagnosis
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Neuroblastoma / genetics*
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Neuroblastoma / mortality
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Neuroblastoma / pathology
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Neurons / metabolism*
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Neurons / pathology
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Poly (ADP-Ribose) Polymerase-1 / genetics
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Poly (ADP-Ribose) Polymerase-1 / metabolism
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Prognosis
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RNA, Long Noncoding / antagonists & inhibitors
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RNA, Long Noncoding / genetics*
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RNA, Long Noncoding / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Signal Transduction
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Sister Chromatid Exchange
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Survival Analysis
Substances
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CENPA protein, human
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Cell Cycle Proteins
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Centromere Protein A
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Chondroitin Sulfate Proteoglycans
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Chromosomal Proteins, Non-Histone
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MYCN protein, human
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N-Myc Proto-Oncogene Protein
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NORAD long non-coding RNA, human
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RNA, Long Noncoding
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RNA, Small Interfering
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SMC3 protein, human
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PARP1 protein, human
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Poly (ADP-Ribose) Polymerase-1
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AURKB protein, human
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Aurora Kinase B
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CDC2 Protein Kinase
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CDK1 protein, human