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BACKGROUND:?Hepatitis?B?virus?(HBV)?is?one?of?the?major?pathogens?of?human?liver?disease.?Studies?have?shown?that?HBV?X?protein?(HBx)?plays?an?important?role?in?promoting?viral?gene?expression?and?replication.?In?this?study?we?performed?a?global?proteomic? profiling? to? identify? the? downstream? functional?proteins?of?HBx,?thereby?detecting?the?mechanisms?of?action?of?HBx?on?virion?replication. METHODS:?HBx? in? the? HepG2.2.15? cell? line? was? knocked?down?by?the?transfection?of?small?interfering?RNA?(siRNA).?The?replication?level?of?HBV?was?evaluated?by?microparticle?enzyme? immunoassay? analysis? of? HBsAg? and? HBeAg? in? the?culture?supernatant,?and?real-time?quantitative?PCR?analysis?of? HBV? DNA.? Two-dimensional? electrophoresis? combined?with?MALDI-TOF/TOF?was?performed?to?analyze?the?changes?in?protein?expression?profile?after?treatment?with?HBx?siRNA. RESULTS:?Knockdown? of? HBx? disturbed? HBV? replication?in vitro.? HBx? target? siRNA? significantly? inhibited? the?expression? of? HBsAg,? HBeAg? and? the? replication? of? HBV?DNA.? Twelve? significantly? changed? proteins? (7? upregulated?and?5?downregulated)?were?successfully?identified?by?MALDI-TOF/TOF? using? proteomics? differential? expression? analysis?after?the?knockdown?of?HBx.?Among?these?identified?proteins,?HSP70?was?validated?by?Western?blotting. CONCLUSION:?The? results? of? the? study? indicated? the?positive? effect? of? HBx? on? HBV? replication,? and? a? group? of?downstream? target?proteins? of?HBx? may? be? responsible?for?this?effect.