Preliminary Study On Synthetic Lethal Effect Of P53-VEGFR2 In Glioma Cells

Purpose: 1.It was preliminarily verified that p53-VEGFR2 has a synthetic lethal effect in human glioma cells.2.To investigate the preliminary effect of p53-VEGFR2 synthesis death in inhibiting the growth of human glioma cells.Research content and methods: The wild-type and mutant p53 human glioma cells U87 and U251 were selected and cultured.First,Western blotting method(WB)was used to verify whether they are wild-type and mutant p53 cells.Si RNAs targeting at three different bands of vascular endothelial growth factor receptor 2(VEGFR2)were designed and synthesized.VEGFR2-siRNA was encapsulated with liposome and transfected into U87 and U251 cells respectively.Each cell is divided into four groups: blank control(BC)does not transfect liposomes,negative control(NC)transfects nonsense sequence-siRNA,Liposome transfection(LT)transfects blank liposomes,and experimental group(VEGFR2-siRNA transfer(VT)transfects VEGFR 2-siRNA of three different bands respectively.The siRNA sequence with the highest inhibition rate and the optimal concentration of the sequence were screened by Western Blotting.The following experiments were carried out using blank control group and experimental group: 1.Western blotting was used to detect the protein expression level of U87 and U251 cells before and after siRNA transfection.2.CCK-8 method was used to detect cell proliferation.3.Scratch test to detect cell mobility;4.Transwell chamber method was used to detect cell invasion.5.Cell aggregation was detected by plate cell clone formation experiment.Results: 1.After siRNA transfection,VEGFR2 protein expression in U87 and U251 cells decreased,and U251 protein expression level was significantly lower than U87.2.CCK-8 detected that U87 and U251 cells were transfected with siRNA,their proliferation ability was weakened,and U251 proliferation inhibition rate was significantly higher than U87.3.Scratch test results show that U87 and U251 have decreased migration ability after siRNA transfection,and U251 migration ability decreases more obviously than U87.4.Transwell cell method showed that the invasive ability of U87 and U251 cells transfected with siRNA decreased significantly compared with U87.5.The experimental results of cell plate cell clone formation showed that the colony forming ability of U87 and U251 cells transfected with siRNA decreased,and the aggregation ability of U251 cells after transfection was significantly lower than U87.Conclusion: 1.p53 and VEGFR2 have synthetic lethal effects in human glioma cells.2.The synthetic lethal of p53-VEGFR2 has obvious inhibitory effect on the growth of human glioma cells.