Expression changes of HIF-1αand its signaling pathway related gene in SH-SY5 Y cells transfected by RIPK3 gene
ZHANG Guolu
CHENG Shixiang
XU Zhongwei
YI Tailong
LIAO Jilian
TU Yue
ZHANG Sai
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.
Keywords：neuroblastomareceptor interacting serine/threonine kinase 3hypoxia-inducible factor 1αubiquitin-conjugating enzyme-like proteinVon Hippel-Lindau proteintranscription elongation factor B polypeptide 1vascular en-dothelial growth factor A
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 13-16 )