Effects Of Antisense Survivin And VEGF Oligodexynucleotides On Human Lung Adenocarcinoma Cell Line A549

ObjectTo investigate the effect of liposome-mediated Survivin and VEGFantisense oligonucleotide used on the growth of human lungadenocarcinoma cell line A549.MethodsThe Survivin and VEGF antisense oligonucleotides wereconstructed,and transfected the human lung adenocarcinoma A549cellswith Lipofectaminel2000. The experiment was divided into control group,liposome group, the the Survivin antisense transfection group (survivinASODN group), the the VEGF antisense transfection group (VEGFASODN group), joint transfection group (Survivin ASODN+VEGFASODN group).The Survivin and VEGF were detected by RT-PCR;FCManalysis was applied to measure cell cycle、cell apoptosis and Survivin andVEGF protein; MTT method was applied to measure cell growth; Colonyforming test was applied to measure the ability of cancer cell to formcolonies. Results1.RT-PCR analysis showed that co-transfection of gene expression ofSurvivin and VEGF lower than the other group, the difference wasstatistically significant (P <0.05).2.Protein expression of Survivin co-transfected group lower than inthe other groups, and the control group was significantly (P=0.017);VEGF expression of co-transfected group lower than in the other groups,and the control group was significantly (P=0.013).3. Cell cycle analysis by flow cytometry showed that co-transfectedcell cycle arrest in S phase was significantly (P <0.05).4. Apoptosis by flow cytometry analysis showed that co-transfectedcell apoptosis was significantly higher than the other group, the differencewas statistically significant (P=0.000, P=0.000, P=0.046, P=0.043).5. MTT assay showed that the combined result of the growth oftransfected cells was significantly lower than the growth capacity of theother groups, the difference was statistically significant (P=0.043, P=0.049, P=0.023, P=0.038).6. Experimental results show that flat colony co-transfected clonal cellproliferation ability was significantly lower than other groups, thedifference was statistically significant (P=0.000, P=0.000, P=0.002, P=0.000). ConclusionsSurvivin and VEGF antisense oligonucleotide combined inhibition ofVEGF gene and protein expression of Survivin, thereby inhibiting thegrowth of human lung adenocarcinoma A549cells, the cells in the S phasearrest，apoptosis rate and The ability of clone increased. For subsequentanimal experiments laid the theoretical basis.