The Study Of Transfection With VEGF Antisense Oligonucleotide On Growth Of Human Gastric Cancer Cells SGC-7901

Objective Our objective is to investigate influence of transfection with VEGF-ASODN on gastric cancer cells SGC-7901.To study elementarily the relationship between VEGF expression and Survivin expression.To observe the change of VEGFmRNA and VEGF protein.In the same time,to observe the change of Survivin protein,apoptosis of the cells and viability of the cells.In the end ,to observe the effect of injection with VEGF-ASODN or Survivin-ASODN on mice's xenograft growth,angiogenesis and VEGF protein in serum.Methods The first part of the experiment focus on gastric cancer cells transfection with VEGF-ASODN,to study the effects of VEGF-ASODN on the growth of SGC-7901.The VEGFmRNA were detected with RT-PCR .The VEGF protein of cells and supernate were detected with ELISA in transfection group and control group respectively.We compared the VEGF protein levels after transfection with VEGF-ASODN or Survivin-ASODN.In the same time ,Survivin protein and apoptosis were detected with Western-blot and FCM.Growth curve of cells was made.Survival was detected by MTT after transfection.Morphologic change was detected by electron microscope. The second part of the experiment is in vivo.We set up control group .We established transplantabal model of SGC-7901 in vivo,and then the mice were put into designed four groups.They were saline group,VEGF group,Survivin group and NOND group.They received injection of saline,VEGF-ASODN,Survivin-ASODN or NODN .We observed growth of nude .The volume and the weight of xenografts were measured.Angiogenesis of tumors were detected by immunohistochemistry.We detected VEGF protein in serum.The expression of PCNA was detected with FCM.Result After the gastric cancer cells SGC-7901 had been transfected with VEGF-ASODN,the VEGFmRNA copies declined dependent on concentration,the amount of VEGF protein in gastric cancer cells and supernate declined dependent on concentration .The Survivin protein declined dependent on concentration too.The VEGF protein in supernate declined after transfection with VEGF-ASODN or Survivin-ASODN .The growth of cells were suppressed.Survival of cells declined and apoptosis of cells increased.Apoptosis change appeard under electron microscope.After received injection of VEGF-ASODN or Survivin-ASODN,we observed decline of angiogenesis and growth of tumor. VEGF proteins in serum,weight and volum of the xenografts decreased.The expression of PCNA decreased after treated with VEGF-ASODN or Survivin-ASODN on xenografts.Conclusion Transfection with VEGF-ASODN on gastric cancer cells SGC-7901 can not only decrease copies of VEGFmRNA,expression of VEGF protein and suppress blood vessel growth of xenografts,but also can decrease expression of Survivin protein and cell's viability. It increases cell's apoptosis.It can decline the level of VEGF protein in vivo too. The growth of xenografts was suppressed. Transfection with Survivin-ASODN on gastric cancer cells SGC-7901 can decrease expression of VEGF protein and the level of VEGF protein in vivo.It can also suppress angiogenesis and growth of xenografts. As a result,antisense therapy can be used for gastric cancer.