Objective:To observe the influence of lentiviral vector-mediated RNA interference on expression of CTGF(connective tissue growth factor,CTGF)gene in human hepatocellular carcinoma cell line. Methods:The effective RNA interference sequence targeting CTGF gene was screened and determined according to our previous study. The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was desiged,synthesized and cloned into Plk0.1-GFP-SP6 vector to construct a lentiviral vector which expressed CTGF shRNA,subsequently confirmed by PCR and DNA sequencing analysis. 293T cells were co-transfected with the plasmids using liposome transfection methods,and packaged to produce lentiviral particles. The recombinant lentiviruses were transfected into HepG2 cells,and the CTGF mRNA and protein expression were examined by Real-time PCR and Western-blot. The results were compared with those of the non-transfected and blank vector transfected HepG2 cells. Results:PCR analysis and DNA sequencing confirmed that the CTGF shRNA sequences were successfully inserted into the lentiviral vectors. After transfection with CTGF shRNA,the CTGF expression in HepG2 cells was significantly inhibited at both mRNA and protein levels compared with that in non-transfected and empty vector transfected HepG2 cells. Conclusion:Two lentiviral RNAi vectors targeting CTGF gene have been successfully constructed,and can effectively inhibit the expression of CTGF gene in HepG2 cells. It paves a way for CTGF-targeted gene therapy of human hepatocellular carcinoma.