Objective To approach whether over-expressed p62 could aggregate more ubiquitinated proteins in DRibbles. Methods Recombinant pIRES2-EGFP vector containing the HBsAg or UbHBsAg gene was constructed and transfected into HepG2 cell line respectively (group HBsAg and group Ub-HBsAg) and pIRES2-EGFP-UbHBsAg and pIRES2-EGFP-p62 were co-transfected into HepG2 cell line (group Ub-HBsAg + p62). Then the transfected cells were cultured in the medium including Velcade, Rapamycin and ammonium chloride. After 16 hours, we extracted and lysed DRibbles from the medium and detected HBsAg by ELISA. Results The level of HBsAg in the DRIbbles extracted from HepG2 cells tranfected with pIRES2-EGFP-Ub-HBsAg was much lower than group HBsAg, while there was no significant difference between group Ub-HBsAg and Ub-HBsAg + p62. However, both of HBsAg levels in group Ub-HBsAg and group Ub-HBsAg + p62 increased rapidly when the cells were cultured with the drugs, and the latter was much higher than the former. Conclusion Over-expressing of p62 aggregated more ubiquitinated proteins (HBsAg) in the DRibbles extracted from HepG2 cells as the proteasome pathway was blocking by the drugs.