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1、HBx及其不同突变体对Hippo信号途径表达的影响陈琰琰1朱慧芬3杨道锋4刘晓聪1邓科兰4黄春梅3胡晓萌4姜素华23沈关心3(832002新疆维吾尔族自治区石河子石河子大学医学院：生物化学教研室1，病原生物学与免疫学教研室2；430030湖北武汉华中科技大学同济医学院免疫系3华中科技大学同济医学院附属同济医院感染科4）［提要］目的初步探寻HBx及其不同突变体表达与Hippo信号途径的关系，以及对细胞凋亡的影响。方法：HBx及其突变体载体转染人正常肝细胞系L02，转染48h后，提取总蛋白，Westernblot
2、检测细胞中Hippo信号途径Mst1/2、YAP(Yes-associatedprotein)的表达，磷酸化和去磷酸化的情况。采用AnnexinV/PI标记流式细胞术检测细胞凋亡。结果：HBx及其突变体载体转染L02细胞48h后，Westernblot结果显示细胞MST1/2表达下调，P-MST1/2表达上升；YAP表达上升，P-YAP表达下调。AnnexinV/PI标记法流式细胞术结果显示HBx及其突变体能促进L02细胞发生凋亡，其介导的细胞早期凋亡尤为明显。结论：HBx通过hippo信号通路途径调控下游致
3、癌基因YAP的表达。结合HBx介导的L02细胞凋亡这一结果说明HBx是一类多功能调节蛋白，可能通过多种途径调节细胞周期。[关键词]：hippoYAP乙型肝炎病毒X蛋白乙型肝炎病毒肝癌细胞凋亡TheeffectsofHBxandit’smutantonHipposignalingpathwayandcellapoptosisinahumanhepaticcellline1ChenYanyan,3ZhuHuifen,4YangDaofeng,1LiuXiaocong,4DengKelan,3HuangChunme
4、i,4HuXiaomeng,2JiangSuhua,3ShenGuanxin（1DepartmentofBiochemicalandMolecularBiology，2DepartmentofImmunology，MedicalCollegeofShiheziUniversity;Shihezi,XinjiangUygurAutonomousRegion,832003;3DepartmentofImmunology，4Departmentofinfection,TongjiHospital;TongjiMed
5、icalCollege;HUST;Wuhan;HubeiProvince;430030）［Abstract］ObjectiveToexploretherelationshipofHBxanditsmutanttoHipposignalingpathwayandtheeffectsofHBxoncellcycleandapoptosis.MethodsL02cellsweretransfectedwithHBxanditsmutant.After48hofincubation,totalproteinwasob
6、tainedandtheexpressionlevelsaswellasthephosphorylationstatusofMst1/2andYAPweredetectedbyWesternblot.TheapoptosisofL02cellwasexaminedcytofluorometricallybyAnnexinV/PIstaining.ResultsAftertransfection,theresultsofWesternblotshowedthattheexpressionlevelofP-MST
7、1/2andYAPwereup-regulatedandthelevelofMST1/2andP-YAPweredown-regulated,Intheresultsofflowcytometry,moreapoptosiscellswereobservedinexperimentalgroupcomparedwiththecontrolgroup,thetrendisparticularlyapparentforearlyapoptosis.ConclusionHBxcouldregulatetheexpr
8、essionofYAPthroughHipposignalingpathway.ThismechanismmayrelevanttoHBVrelatedHCC,combinedwiththeresultsofflowcytometry,anditsuggestedthatHBxisamultifunctionalproteinthatregulatesthecellcycleinseveralway
