Abstract: Objective：To construct a lentiviral vector of homo sapiensmiR-152 (hsa-miR-152) and screen the cell strain for its stable expression. Methods：The fragments of pre-miR-152 were cloned from genomic DNA of human adipose tissue and inserted into lentiviral vector pGIPZ. The recombinant plasmid pGIPZ-miR-152 was confirmed by restriction endonuclease analysis and DNA sequencing. 293T cells were transfected with the recombinant plasmid pGIPZ-miR-152 and the supernatant containing the lentivirus particles was harvested to determine the virus titer and used to infect HepG2 cells. Puromycin was used to screen the infected cells, so as to obtain a cell strain for stable expression. The expression of miR-152 was determined using real-time PCR. Results：The restriction enzyme digestion and DNA sequencing demonstrated the lentiviral vector pGIPZ-miR-152 was constructed. The lentiviral vector was transfected into 293T cells and the viral titer reached 7.5×10⁷ TU/mL. HepG2 cells were infected by the lentivirus and the infection efficiency was over 70%. The expression of miR-152 was increased nearly 10 folds in the overexpression group following screening with puromycin for 10 days. Conclusion: The lentiviral vector expressing hsp-miR-152 was constructed successfully. The cell strain with stable overexpression of miR-152 was also obtained, which provided the material for the further study of molecular function of miR152 in nonalcoholic fatty liver disease.