Résumé
Objective To observe the effects of VEGF antisense oligonucleotide transfected to SGC-7091 cell line on expression of VEGF and survivin of the cell.Methods The VEGF-ASODN was synthesised artificially.There were five groups in this experiment: control group,missense group and three various concentration groups.After transfection,RNA copies were detected by real-time PT-PCR,VEGF protein in cell and culture supernate was detected by ELESA,survivin protein was measured by Western-blot,and apoptosis was detected by FCM.Growth of cells was evaluated by growth curve.ResultsVEGF-ASODN tranfection reduced the VEGFmRNA and VEGF protein expression in gastric cancer cells and supernate remarkably.It also reduced survivin protein expression and increased apoptosis in gastric cancer cells.Conclusions Transfection with VEGF-ASODN to gastric cancer cells SGC-7901 can reduce the expression of VEGF,and can increase apoptosis and suppress the proliferation of gastric cancer cells.
Résumé
Objective To explore the suppressive effect of survivin antisense oligonucleotide on transplanted human breast cancer in nude mice. Methods 30 nude mice were transplanted with human breast cancer MCF-7 cells to construct tumor models, which were divided into three groups (10 mice per group): (Lipofectin), ASODN/Lip, and NODN/Lip were injected in to the tumor or around the tumor, respectively. A total of 3 injections were given, once every five days. Observations were made on tumor suppression rate, tumor volume and changes in weight of the animals. TUNEL method and transmission electron microscopy were used to investigate cell apoptosis of the tumor; western blot was used to detemine the expression of survivin protein in the 3 groups. Results The subcutaneous tumor model in nude mice was successfully established. The tumor volume in the group of ASODN/Lip decreased during treatment, the rate of tumor suppression reached 70.10%; while the tumor volume of the other two groups increased. There was no statistical (difference) among the groups in weight changes of the 3 groups mice, the apoptosis rate in ASOND/Lip group was 38.6% detected by TUNEL and the rate was markedly higher than the other two groups(P
Résumé
Objective To observe the effect of PKB gene silencing on the growth of gastric cancer cell line SGC-7901 in vitro.Methods Gene transfection technique was used to transfect AKt2 siRNA into gastric cancer cells.Akt2 expression was detected by RNAi technique,Akt2 protein level was detected by Western blot,and the change of cell cycle distribution and apoptosis of SGC-7901 cells were detected by flow-cytometry,SGC-7901 proliferation was measured by MTT method.Results After SGC-7901 cells transfected with Akt2 siRNA,the expression of protein level decreased obviously(P