〔Abstract〕 Objective To construct lentiviral vector ofp62gene over-expression, and stably expressp62gene in human monocytic leukemia cells 1(THP-1), and to provide a way to study the role ofp62gene at the cellular level. Methods Thep62gene fragment was amplified by polymerase chain reaction(PCR), and the amplified product was ligated to the linearized pcDNA3.1-Flag-PCDH10 lentiviral vector. After identifying with PCR, the PCR product was cotransfected with the packaging plasmid into human embryonic kidney cells 293(HEK 293T). THP-1 cells were infected with recombinant lentivirus. Positive cell clones were screened by ampicillin. Western blot and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR) were used to detect THP-1 cell lines with highp62expression(overexpression group) and THP-1 cell lines transfected with empty plasmid withoutp62gene(control group). The expression levels of TNF-α, IL-1β and Cxcl1 afterK.p. infection were detected by RT-qPCR. Results Thep62gene fragment was successfully obtained by PCR and ligated to pcDNA3.1-Flag-PCDH10 vector. PCR confirmed thatp62-pcDNA3.1-Flag-PCDH10 recombinant plasmid was constructed successfully. Ampicillin-resistant cell lines were selected after lentiviral infection of THP-1 cells. The results of Western blot analysis showed that the THP-1 cells with drug sieve survival increased the expression of P62 protein compared with the control cells(P<0.001), and RT-qPCR analysis showed that the relative mRNA expression ofp62increased(P<0.001). THP-1 cells with high expression of P62 were successfully constructed. The levels of TNF-α、IL-1β and Cxcl1 from THP-1 cells with high expression of P62 significantly increased after infection withK.p.(P<0.01). Conclusion P62-pcDNA3.1-Flag-PCDH10 vector and THP-1 cells with high expression of P62 can be successfully constructed by three-plasmid packaging system, which provides a basis for the study ofp62.