The full length sequence of GLI1 gene was amplified by PCR method and it was cloned into the pcDNA3.1 eukaryotic expression vector. The PC9 cells were transfected with the recombinant vector pcDNA3.1-GLI1 （recombination vector group） and the control vector pcDNA3.1 （empty vector group）. The construction of GLI1 over-expression vector was detected by enzymatic digestion identification method. The cells stably expressed with GLI1 and empty vector were established by G418 screening. The expression levels of GLI1 mRNA in the PC9 cells in two groups were detected by RT-PCR method； the expression levels of GLI1 protein in the PC9 cells in two groups were detected by Western blotting method；the proliferation activities of the PC9 cells in two groups were detected by CCK-8 method； the clone formation number of the PC9 cells in two groups was detected by cell clonal formation assay.

The GLI1 gene fragment was specifically amplified successfully； the target genes and vector fragments were obtained by double enzyme digestion of the GLI1 eukaryotic expression vectors. The RT-PCR results showed that the expression level of GLI1 mRNA in the PC9 cells in recombination vector group was significantly higher than that in empty vector group （P<0.01）. The Western blotting results showed that the expression level of GLI1 protein in the PC9 cells in recombination vector group was significantly higher than that in empty vector group （P<0.05）. The CCK-8 results showed that the proliferation activities of the PC9 cells in recombination group were significantly higher than those in empty vector group at 72 and 96 h after culture （P<0.01）. The clonal formation assay results showed that the clone formation number of the PC9 cells in recombination group was significantly more than that in empty vector group at 10 d of culture （P<0.01）.

The GLI1 eukaryotic over-expression vector and the PC9 cells stably over-expressed with GLI1 are established successfully. Over-expression of GLI1 can promote the proliferation ability of lung cancer PC9 cells.