Objective：To study the effects of Pellino1’s post-translational modification—small ubiquitin-related modifier（SUMO）modification of Pellino1 in the nuclear factor κB（NF-κB）signaling pathway mediated by the TNF receptor associated factor6（TRAF6）. Methods：We constructed the plasmid of the SUMO modification sites mutantation of Pellino1，then cotransfected with TRAF6 plasmid and ubiquitin（Ub）plasmid into HEK-293 cells. The efficiency of plasmid transfection was observed by fluorescence microscopy. We detected the combination of mutant Pellino1 with TRAF6 by co-immunoprecipitation，and monitored the ubiquitination level of Pellino1 and TRAF6. Lipopolysaccharide（LPS）was used to induce inflammatory response，and the cytoplasmic nucleocapsid was extracted. Western blot was used to detect the phosphorylation of inhibitor of NF-κBα（IκBα）and the nuclear translocation of NF-κB P65. The effect of Pellino1 SUMO modification on TRAF6 mediated NF-κB signaling pathway was analyzed. Results：Compared with wild type Pellino1，mutant of Pellino1 SUMOylation ylation not only increased ubiquitination of Pellino1 itself，but also the combination with TRAF6，and the ubiquitination modification of TRAF6 were also increased. LPS stimulation significantly increased the phosphorylation of inhibitor of NF-κBα（IkBα）and NF-κB P65 nuclear translocation. High expression of mutant of Pellino1 SUMO after transfection significantly enhanced the LPS induced NF-κB signal activation. Conclusion：Mutant of Pellino1 SUMOylation can promote TRAF6 mediated activation of NF-κB signaling pathway by increasing the ubiquitination of Pellino1 and binding with TRAF6 and the ubiquitination level of TRAF6.