Abstract: Objective To evaluate the effects of MMP-9 gene down-regulation by antisense on proliferation of a human glioblastoma cell line. Methods A 528bp cDNA fragment of MMP-9 was amplified by RT-PCR with synthetic primers and subcloned into the pcDNA3.0 vector in the sense and antisense orientations. Then TJ905 cells were transfected with sense construct, antisense construct and pcDNA3.0 by using Lipofectamine 2000. MMP-9 RNA and protein were detected by RT-PCR and Western blot respectively. MMP-9 and Ki-67 expression were detected by immunohistochemistry assay. Results Restriction endonuclease analysis and sequence analysis of recombination plasmids verified that 528bp internal sequence was 100% homology with the published sequence of MMP-9 cDNA. Compared with controls, vector-transfected clones and sense MMP-9 construct-transfected clones, antisense MMP-9 construct-transfected TJ905 cells had decreased MMP-9 mRNA and protein levels(P<0.001). The proliferation of antisense MMP-9 construct-transfected TJ905 cells was inhibited. No significant difference in expression of MMP-9 was observed among vector-transfected clones, sense MMP-9 construct-transfected clones and controls（P>0.05）. Conclusion Proliferation of glioblastoma cell was able to be effectively inhibited through antisense MMP-9, suggesting antisense MMP-9 may be useful for malignant glioma treatment in future.