Survivin Asodn Combined With VEGF Asodn Inhibits The Growth Of Human Pulmonary Adenocarcinoma A549Xenografts In Nude Mice

Objective： The aim of this study was to investigate the inhibitoryeffect of Survivin antisense oligonucleotide combined with VEGF antisenseoligonucleotide on the human pulmonary adenocarcinoma A549xenograftsin nude mice.Methods： Take a certain amount of normal A549cells oflogarithmic growth phase injected into nude mice.Then, the25A549xenograft nude mice models were randomly divided into5groups (eachgroup with5nude mice): control group，lipofectamine control group,Survivin treatment group, VEGF treatment group, Survivin-VEGFcombined treatment group. Those groups was dealed8times withphysiological saline, lipofectamine, Survivin ASODN, VEGF ASODN,Survivin ASODN﹢VEGF ASODN, respectively.After those,we observedtumor growth of each groups and used western blot and rtPCR to evaluatedthe expressions of the targeted genes in tumor tissues. Usingimmunohistochemical method to evaluated the expressions of Caspase3and Cyclin B1,and to count the microvascular density (CD34markers).UsingTUNEL staining method to detect the apoptosis in the tumor tissues.Results： VEGF antisense oligonucleotide and Survivin antisenseoligonucleotides, whether combined or alone, can effectively downregulated the expression of target genes and inhibit the growth of the tumor,and the combined treatment group is better than single antisenseoligonucleotide treatment groups. The Tumor weight of Survivin treatmentgroup is (0.479+0.046) g, The Tumor weight of VEGF treatment group is(0.476+0.079) g, The Tumor weight of of Survivin-VEGF combinedtreatment group is (0.265+0.010) g, there is a significant differencebetween those groups with the blank group (P <0.05), and the combinedtreatment group is better than the single antisense oligonucleotide treatmentgroups(P <0.05). The protein and RNA expressions of Survivin and VEGFin Survivin-VEGF combined treatment group is lower than the Survivintreatment group and VEGF treatment group(P<0.05). Moreover, Survivin-VEGF combined treatment group significant decreased the counting ofmicrovascular density (MVD) in tumor tissues more than the singleantisense oligonucleotide treatment group, Survivin treatment group (13.27+2.19) and VEGF treatment group (11.40+1.18), Survivin-VEGFcombined treatment group (10.27+1.75), there is a significance differencebetween single antisense oligonucleotide groups with combined group (P <0.05). Survivin-VEGF combined treatment group retardant more cells in the G2/M phase,there are also a significantly difference between theSurvivin-VEGF combined treatment group with the Survivin treatmentgroup and the VEGF group (P <0.05).Conclusion： The combination of VEGF antisense oligonucleotideand Survivin oligonucleotide can get a better inhibition of the tumorgrowth than using VEGF antisense oligonucleotides or Survivin antisenseoligonucleotides alone.