Yang Chunhui, Qu Jiaying, Cheng Yang, Tian Minxiu, Wang Zhijie, Wang Xiaolin, Li Xinyue, Zhou Shunchen, Zhao Bosen, Guo Yanhua, Zheng Liduan, Tong Qiangsong
Department of Pediatric Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, 430022, Hubei Province, People's Republic of China.
Department of Pathology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, 430022, Hubei Province, People's Republic of China.
J Transl Med. 2024 Dec 27;22(1):1153. doi: 10.1186/s12967-024-05956-4.
Neuroblastoma (NB), the most prevalent solid tumor in children, arises from sympathetic nervous system and accounts for 15% of pediatric cancer mortality. This malignancy exhibits substantial genetic and clinical heterogeneity, thus complicating treatment strategies. Poly(ADP-ribose) polymerase 1 (PARP1), a key enzyme catalyzing polyADP-ribosylation (PARylation), plays critical roles in various cellular processes, and contributes to tumorigenesis and aggressiveness. However, the functions and regulatory mechanisms of PARP1 in NB progression still remain to be determined.
The association of PARP1 expression with NB patients' survival was analyzed by mining of R2 database. Western blotting, reverse transcription-polymerase chain reaction, MTT colorimetric, soft agar, and matrigel invasion assays were utilized to assess PARP1 expression and its effects on aggressiveness of NB cell lines. Chromatin immunoprecipitation (ChIP) sequencing and ChIP assays were employed to investigate the binding of Yin Yang 1 (YY1) to PARP1 promoter. Protein interactions were explored by BioGRID database analysis, molecular docking, and co-immunoprecipitation assay. RNA sequencing and crosslinking-immunoprecipitation high throughput sequencing datasets were used to identify precursor mRNA splicing targets of non-POU domain containing octamer binding protein (NONO).
High PARP1 expression was associated with poor survival of NB patients. PARP1 over-expression enhanced the proliferation and invasion of NB cell lines, confirming its oncogenic roles. YY1 was identified as a key transcriptional regulator facilitating PARP1 expression. Additionally, PARP1 interacted with NONO to induce its PARylation, resulting in stabilization of NONO protein via preventing ubiquitin-mediated degradation. NONO facilitated the splicing and mRNA maturation of target genes a disintegrin and metalloproteinase domain 8 (ADAM8) and testis-expressed gene 14 (TEX14) in a PARylation-dependent manner. Rescue experiments indicated that YY1 facilitated PARP1-mediated PARylation of NONO and subsequent mRNA maturation of ADAM8 and TEX14 in NB cells. In clinical NB cases, high expression of YY1, PARP1, NONO, ADAM8, or TEX14 was associated with poor survival of patients.
These findings indicate that YY1 drives PARP1 expression essential for PARylation of NONO in mRNA maturation during NB progression.
神经母细胞瘤（NB）是儿童中最常见的实体瘤，起源于交感神经系统，占儿童癌症死亡率的15%。这种恶性肿瘤表现出显著的遗传和临床异质性，从而使治疗策略复杂化。聚（ADP-核糖）聚合酶1（PARP1）是催化聚ADP-核糖基化（PARylation）的关键酶，在各种细胞过程中发挥关键作用，并促进肿瘤发生和侵袭性。然而，PARP1在NB进展中的功能和调控机制仍有待确定。
通过挖掘R2数据库分析PARP1表达与NB患者生存的相关性。采用蛋白质印迹法、逆转录-聚合酶链反应、MTT比色法、软琼脂法和基质胶侵袭试验评估PARP1表达及其对NB细胞系侵袭性的影响。采用染色质免疫沉淀（ChIP）测序和ChIP试验研究阴阳1（YY1）与PARP1启动子的结合。通过BioGRID数据库分析、分子对接和免疫共沉淀试验探索蛋白质相互作用。利用RNA测序和交联免疫沉淀高通量测序数据集鉴定含八聚体结合蛋白（NONO）的非POU结构域的前体mRNA剪接靶点。
PARP1高表达与NB患者的不良生存相关。PARP1过表达增强了NB细胞系的增殖和侵袭，证实了其致癌作用。YY1被鉴定为促进PARP1表达的关键转录调节因子。此外，PARP1与NONO相互作用以诱导其PARylation，通过防止泛素介导的降解导致NONO蛋白稳定。NONO以PARylation依赖的方式促进靶基因解整合素和金属蛋白酶结构域8（ADAM8）和睾丸表达基因14（TEX14）的剪接和mRNA成熟。挽救实验表明，YY1促进PARP1介导的NONO的PARylation以及随后NB细胞中ADAM8和TEX14的mRNA成熟。在临床NB病例中，YY1、PARP1、NONO、ADAM8或TEX14的高表达与患者的不良生存相关。
这些发现表明，YY1驱动PARP1表达，这对于NB进展过程中mRNA成熟过程中NONO的PARylation至关重要。
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