山东医药Issue(7)：13-16,4.DOI:10.3969/j.issn.1002-266X.2016.07.005
RIPK3基因转染的 SH-SY5Y 细胞中 HIF-1α基因及其信号通路相关基因表达变化
Expression changes of HIF-1αand its signaling pathway related gene in SH-SY5 Y cells transfected by RIPK3 gene
摘要
Abstract
Objective To observe the expression changes of hypoxia-inducible factor 1α( HIF-1α) and its signaling pathway related gene in the neuroblastoma cell line SH-SY5Y transfected by receptor interacting serine/threonine kinase 3 (RIPK3) gene.Methods The pCMV6-AC-GFP plasmid expressing RIPK3 gene (recombinant plasmid) was construc-ted.SH-SY5Y cells were cultured and then were transfected with the recombinant plasmid and empty vector plasmid as the experimental group and control group respectively.Expression of RIPK3 protein was detected by Western blotting, and the OD value was detected by MTT assay at 8, 14, 20, 26, 32 and 38 h.RNA transcriptome sequencing (RNAseq) and Inge-nuity Pathway Analysis ( IPA) was used to detect and screen the key genes in the downstream signaling pathway of RIPK3-HIF-1α.Droplet Digital PCR ( ddPCR) was used to detect the HIF-1αmRNA in the two groups.Results The RIPK3 ex-pression in the experimental group (0.806 ±0.097 5) was higher than that of the control group (0.455 ±0.088 6), P<0.05.The proliferation of SH-SY5Y was inhibited as the cell incubation time was prolonged.The expression of HIF-1αmRNA in the experimental group (0.01543 ±0.00347) was strongly down-regulated as compared with that of the control group (0.04628 ±0.01026) ( P<0.05) .In the interaction network of HIF-1αwhich was taken as the core, we screened the key molecules such as ubiquitin-conjugating enzyme ( UBC) , von Hippel-Lindau ( VHL) , transcription elongation fac-tor B polypeptide 1 (TCEB1) and vascular endothelial growth factor A (VEGFA).Conclusion After the SY5Y cells were trasfected by RIPK3, the HIF-1αmRNA expression was down-regulated in SH, and the expression levels of HIF-1αsignaling pathway-related genes ( UBC, VHL, TCEB1 and VEGFA) were affected.关键词
神经母细胞瘤/受体相互作用蛋白激酶3/低氧诱导因子1α/泛素缀合酶样蛋白/希佩尔-林道蛋白/转录延伸因子B多肽/血管内皮生长因子AKey words
neuroblastoma/receptor interacting serine/threonine kinase 3/hypoxia-inducible factor 1α/ubiquitin-conjugating enzyme-like protein/Von Hippel-Lindau protein/transcription elongation factor B polypeptide 1/vascular en-dothelial growth factor A分类
医药卫生引用本文复制引用
张国禄,程世翔,徐忠伟,衣泰龙,廖吉连,涂悦,张赛..RIPK3基因转染的 SH-SY5Y 细胞中 HIF-1α基因及其信号通路相关基因表达变化[J].山东医药,2016,(7):13-16,4.基金项目
国家自然科学基金项目（31200809）；武警部队后勤科研项目（WJHQ2012-20）。 （）
