DExH-box解旋酶9O-GlcNAc修饰对HBV相关肝癌细胞增殖能力的影响
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【中图分类号】R735.7 【文献标志码】A
Effect of DExH-box helicase 9 O-GlcNAc modification on the proliferation of HBV-associated hepatoma cells
LuoHuimin，PiYubo，Chen Yanmeng，WangKai，TangNi，HuangAilong （KeyLaboratory ofMolecular Biology on Infectious Diseases， MinistryofEducation，ChongqingMedical University）
【AbstractObjective：ToinvestigatetheeffectofDExH-boxhelicase9（DHX9）O-linkedN-acetylglucosamine（O-GlcNc）modifcation（O-GlcNAcylation）ontheproliferationofhepatitisBvirus（HBV）-associatedhepatomacels.Methods：ThepAdTrack-TO4- DHX9-3Flagrecombinantadenovirus plasmid wasconstructedbymolecularcloningand transfectedinto HEK293cellsfrpackaging andamplificationoftherecombinantadenovirusAdDHX9.TheinteractionbetweenDHX9andO-GleNActransferase（OGT）wasconfirmedusingco-immunoprecipitation.Theco-lcalizationbetweenDHX9andOGTwas measuredbyimmunofluorescence.Thelevel ofDHX9O-GlcNAcylationwasdetermnedusingsucinylatedwheat germaglutinin（sWGA）andglcosylatedimmunoprcipitation （IP）.The efectof HX9O-GleNAcylationontheproliferationofHBV-associatedhepatomacellswasassedusingthecolony forming assayandcellgowthurves.Results：Therecombinant adenovirus dDHX9wassuccesfullyobtaied，andHX9expsion wasconfirmedbyWesternblot.DHX9 interactedwithOGT，andthetwoproteinswereco-localizedonthenucleus.ThesWGAandglycosylated PexperimentsshowedthatDHX9underwentO-GlcNAcylation，whichwasfurtherenhancedbyHBVinfection.Thecolonyforming assay demonstrated that the number of cell clones was increased in the AdDHX9 g roup（386.667±15.630） compared with the AdGFP control group（142.667 ± 7.572， P<0.001 ）.Moreover，cell growth curves demonstrated that the overall cell growth rate was enhancedinthe AdDHX9 group（22.860±0.770） compared with the AdGFP control group（13.670±0.517，P<0.001）.Conclusion：HBV infection promotes DHX9O-GlcNAcylation，which enhancesthe proliferation of HBV-associated hepatoma cells.
【Key words]DExH-box helicase9;O-linked N-acetylglucosamine; hepatocellular carcinoma;cell proliferation
原发性肝癌是全球常见的恶性肿瘤之一，居恶性肿瘤发病率的第6位，其中肝细胞癌（hepatocellularcarcinoma，HCC）占 75%～85%[1] 。（剩余16697字）