关键词: 神经母细胞瘤, 细胞系, 肿瘤, 锌指, 基因表达调控, 血管内皮生长因子类, 脑源性神经营养因子, 肿瘤坏死因子类, 受体相互作用蛋白激酶-3, 锌指蛋白36, 人脱帽酶2

Abstract: Abstract: Objective To investigate the signaling pathway and the key signal molecules of protein kinase (RIPK)3 in SH- SY5Y cells. Methods SH-SY5Y cells were transfected with RIPK3 expression plasmid vector to upregulate intracellular RIPK3, while the SH-SY5Y cells were transfected with empty vector plasmid, which was considered as control group. Western blot assay was used to check the expression of exogenous RIPK3 in cells. The proliferation rate of SH-SY5Y cells was determined by MTT assay at designated time to detect exogenous RIPK3 activity. Whole transcriptome sequencing (RNAseq) was used to detect the transcription of genes. Whole-transcriptomic gene transcription was measured by following Ingenuity Pathway Analysis (IPA) to obtain downstream signaling pathways and the key molecule, which were partly confirmed by following droplet digital PCR (ddPCR). Results Exogenous RIPK3 showed biological activity in SH-SY5Y, which inhibited the proliferation of cells. IPA showed that znic finger protein 36 (ZFP36) was significantly up-regulated as compared with that of the control group. The tran⁃ scription levels of ZFP36 downstream genes such as tumor necrosis factor (TNF), brain derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF) and mRNA-decapping enzyme 2 (DCP2) were affected at the same time. Conclusion Within the limitations of this study, it seems that RIPK3 is notable for the development, inflammation and tumorigenesis of the nervous system as an independent regulator of ZFP36 gene and downstream effectors.

Key words: neuroblastoma, cell line, tumor, zinc fingers, gene expression regulation, vascular endothelial growth factors, brain-derived neurotrophic factor, tumor necrosis factors, receptor-interacting protein kinase 3, znic finger protein 36, mRNA-decapping enzyme 2