〔Abstract〕 Objective To construct a mouse derived pcDNA3.1-3 × Flag-c-NUP85 expression plasmid and observe its effect on expression of inflammation factors in LPS-induced RAW264.7 cells,as well as on the proliferation and apoptosis of RAW264.7 cells.Methods The NUP85 gene was amplified by PCR to construct pcDNA3.1-3 × Flagc-NUP85 eukaryotic expression plasmid.The pcDNA3.1-3 × Flag-c vector was divided with enzymes.The purified PCR product was ligated with the vector,and the ligated product was transformed into bacterial competent cells.After identification by enzyme digestion,sequencing and analysis were performed.Then,it was transfected into RAW264.7 cells,and the blank plasmid without NUP85 gene was set as the control group.The effect on cell proliferation and apoptosis were detected by CCK-8 assay and flow cytometry,and the expression of inflammatory cytokines such as tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in LPS-induced RAW264.7 cells was detected by Western blot and ELISA.Results Enzyme digestion identification and Western blot results showed that pcDNA3.1-3 × Flag-c-NUP85 eukaryotic expression plasmid was successfully constructed and expressed.The results of CCK-8 assay showed that the cell survival rate of NUP85 overexpression group was significantly lower than that of control group after 24 h [(0.55±0.03) vs(0.67±0.05),F=30.98,P<0.05].The results of flow cytometry showed that the cell apoptosis rate of NUP85 overexpression group was higher than that of control group [(15.78±1.05) % vs(13.40±0.47) %,F=75.38,P<0.05].The results of Western blot and ELISA showed that after transfection of pcDNA3.1-3 × Flag-c-NUP85,the expression of TNF-α and IL-6 in RAW264.7 cells were higher than those in the control group,with statistical significance(P<0.05).Conclusion NUP85 can inhibit the proliferation and promote apoptosis in LPS-stimulated RAW264.7 cells,and NUP85 can promote the expression of inflammatory cytokines IL-6 and TNF-α in LPS-stimulated RAW264.7 cells.