Abstract: [
Abstract ] Objective Theaimofthisstudyistoinvestigatethedownstream pathwayof
receptor-interactingproteinkinase3 ( RIPK3 ) .Methods SH-SY5Y wasstablytransfectedwith
pCMV6-RIPK3recombinantplasmidtoup-regulateexpressionofRIPK3 , whilecontrolgroup
wastransfectedwithemptyvector.RIPK3levelintransfectedSH-SY5Ycellsweredetermined
usingwesternblot.Cellproliferationactivitywasmeasuredby MTTtodetecttheactivityof exogenousRIPK3.Thedifferentiallytranscriptedgenesbetweentwogroupswereidentifiedby
RNAseq.Ingenuitypathwayanalysis ( IPA ) revealedagreatroleofRIPK3anditsdownstream
signalpathwaysandthekeymoleculemammaliantargetofrapamycin ( mTOR ) .Dropletdigital
PCR ( ddPCR ) analysiswasusedtofurtherconfirmexpressionofmTOR.Results PlasmidDNA
wasintegratedintothecellgenome.WesternblottingresultssuggestedthatexogenousRIPK3
wasexpressedinexperimentalgroup.MTTresultsshowedthatexogenousRIPK3couldinhibit
cellviability.RNAseqandIPAshownthatoverexpressionofRIPK3upregulatedthetranscription
levelsof mTOR , andaffected downstream genessuch assterol-response binding proteins
(
SREBPs ), p70ribosomalS6proteinkinases ( S6K ), eukaryoticinitiationfactor4E-binding
proteins ( 4E-BPs ) andunc-51-likekinase (
ULK ) .ddPCRanalysisshowthatthechangeofthe
expressionlevelofmTORwasthesameasthatofRNAseq.Conclusion RIPK3couldregulate
transcriptionofmTORsignalcascadeandplayanimportantroleindevelopment , inflammation
andtumorigenesisofthenervoussystem.

Key words: neuroblastoma, receptorinteracting protein kinase 3, gene expression regulation