RESUMO
AIM: Osteosarcoma OS-732 cell line was used to investigate the effect of antisense oligonucleotide (ASODN) on survivin expression and its inducible effect on tumor cells apoptosis. METHODS: ASODN of specific target survivin was designed, synthesized and then transfer to OS-732 cell line with different concentrations and time points. At the same time blank control group, sense oligonucleotide (SOND) group were set up for comparison. Reverse tanscriptionase-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry were used to detect the expressions of survivin mRNA and protein in each OS-732 cell line group. Acridine orange /ethidium bromide (AO/EB) staining and flow cytometry were used to detect the apoptosis level and morphologic change. Mononuclear cell direct cytotoxicity assay (MTT) was used to estimate cell growth suppression. Kinase activity assay method was used to estimate the activity of caspase-3 in the cells. RESULTS: Compared with control group and SOND group, in ASODN groups, the expression of survivin mRNA and protein were obviously weaken, apoptosis rate and caspase-3 activity apparently increased, cells growth was inhibited. In each ASODN group, the effect above-mentioned has time- and concentration-dependent manner. There was no obviously difference of each index in each SODN and blank control groups. CONCLUSION: ASODN down-regulated the expression of survivin gene in OS-732 cell line specifically, and activated apoptosis effectively. It plays an important role in inducing tumor apoptosis and suppressing cell proliferation.