Effects of antisense VEGF and Survivin oligodexynucleotides on human lung adenocarcinoma cell line A549

Abstract:

Objective：To investigate the effect of liposome-mediated vascular endothelial growth factor（VEGF） and Survivin antisense oligonucleotide on the growth of human lung adenocarcinoma cell line A549. Methods:VEGF and Survivin antisense oligonucleotides were constructed and human lung adenocarcinoma A549 cells were transfected with LipofectamineTM2000. The experiment was divided into control group，liposome control group，VEGF antisense transfection group（VEGF ASODN group），Survivin antisense transfection group（Survivin ASODN group），joint transfection group（VEGF ASODN+Survivin ASODN group）. VEGF and Survivin were detected by RT-PCR. Flow cytometry（FCM） was applied to measure cell cycle，cell apoptosis and VEGF and Survivin protein. MTT method was applied to measure cell growth. Colony forming test was applied to measure the cell proliferation and clonogenic abilities. Results：A549 cell viability and proliferation and clonogenic abilities were significantly lower in VEGF ASODN+Survivin ASODN group than in the other groups，with significantly statistical differences（P=0.043，P=0.049，P=0.038，P=0.023）. FCM analysis showed that co-trans－fection of transfected cell cycle was arrested and Survivin protein expression was lower in VEGF ASODN+Survivin ASODN group than in the other groups，with significantly statistical differences（P=0.017，P=0.002）. VEGF protein expression was lower in VEGF ASODN+Survivin ASODN group than in the other groups，with significantly statistical differences（P=0.013，P=0.003）. RT-PCR re－sults showed that VEGF and Survivin gene expressions were lower in VEGF ASODN+Survivin ASODN group than in the other groups. Conclusions：Combination of VEGF and Survivn antisense oligonucleotides can inhibit the proliferation of human lung adeno－carcinoma cell line A549，extent cell cycle and inhibit VEGF and Survivin gene expression，which will lay theoretical basis for the subsequent animal experiments.