Abstract:
Objective: To investigate effect of Survivin antisense oligonucleotide(ASODN) on expression of Survivin mRNA and qrotein,and apoptosis in lung cancer cell lines.
Methods: The lung cancer cell lines,NCI-H446 and SPC-A1 were transfected with Survivin ASODN (100 nmol/L,300 nmol/L,500 nmol/L),NSODN(100 nmol/L,300 nmol/L,500 nmol/L),by using Lipofectin for 24 hour,48 hour and 72 hour.The expression of Survivin mRNA was measured by RT-PCR technique.The expression of Survivin protein was measured by western blotting assay,the apoptosis index,proliferation index and cell cycle were measured by flow cytometry (FCM).
Results: Survivin ASODN induced downregulation of the Survivin mRNA of NCI-H446 and SPC-A1 cells in dose-dependent and time-dependent manners,and at concentration of 500 nmol/L for 72 hour,the suppression rate of Survivin mRNA was 62.72% and 67.43%,respectively.Survivin ASODN also induced downregulation of the Survivin protein in both lung cancer cell lines.By analyzing the cell cycle and apoptosis,it was demonstrated that G 0/G 1 and AI were increased and apoptosis was induced with dose-dependent and time-dependent manners.Its maximum effect was achieved at a concentration of 500 nmol/L for 72 hour,at which apoptosis indexes were 44.71% and 31.83%,respectively.
Conclusions: Both NCI-H446 and SPC-A1 express Survivin mRNA and protein;Survivin ASODN can downregulate Survivin mRNA and protein;Survivin ASODN can induce apoptosis in dose-dependent and time-dependent manners,its maxinum effect is achieved at a concentration of 500 nmol/L for 72 hour.