| Objective:Glioma is the most common brain tumor in adults,and its incidence rate accounts for 81 % of intracranial tumors.Since glioma is the largest,highly invasive,and highly malignant tumor,the five-year survival rate of Glioblastoma patients is only 5 %.Although research on glioma has made great progress in recent years,there has been no substantial breakthrough in the treatment of glioma,and the prognosis of glioma is still not optimistic.KDR encodes a protein called Vascular endothelial growth factor receptor-2(vegfr-2),which is an important receptor for VEGF signaling related angiogenic factors.The purpose of this study is to apply the extract of cordycepin to human glioma cell U87,and to explore the inhibition and mechanism of cordycepin on human glioma cell U87,so as to provide new ideas for clinical treatment of human glioma.Methods:Human glioma cell U87 was as the research object,the quantitative effects and aging characteristics of cell growth activity were studied after treatment of cordycepin in different concentrations and different times.The growth inhibition,apoptosis rate,and KDR gene expression of U87 glioma cells were detected after the treatment of cordycepin by flow cytometry,MTT,cell Scratching Experiment and Real-time PCR.The morphology changes of U87 cells were observed at the conditions of 0,20,40,80,160,and 320 μmol/L concentrations of cordycepin.Whereas 0 μmol/L concentration of cordycepin was used as negative control and TMZ was used as positive control.The inhibitory effect and mechanism of U87 cells were researched by scratch experiments,flow cytometry,MTT,and Real-time PCR at different concentrations of cordycepin.Results:1.After treatmented with 0,20,40,80,160,320 μmol/L concentrations of cordycepin,the glioma cell U87 gradually contracted,became round,and had a poor refractive index.As the concentration increased and the time of action prolonged,the phenomenon of cell shedding intensified,and the morphology changed significantly.2.The proliferation of glioma cell U87 was inhibited to varying degrees after the treatmented with 80 mol/L cordycepin for 24 h and 48 h,respectively.3.The results of flow cytometry showed that the apoptosis of U87 cells increased with increasing concentration when 0,20,40,80,160,320 mol/L concentrations of cordycepin was edded,respectively.4.The proliferation of glioma cells U87 was inhibited in a dose-time dependent manner at concentrations of 0,20,40,80,160,320 mol/L and at different periods of time.5.The Real-time PCR results showed that the level of mRNA expression of the KDR gene in the cordycepin group and the TMZ group was significantly lower than that of the control group.With the increase in the concentration of cordycepin,the level of KDR gene expression decreased.Conclusion:1.After the action of cordyceps sinensis,U87 cells showed significant decrease in cell activity and increased cell shedding,with concentration and time dependence;2.After the effect of cordycepin on glioma cell U87,cell proliferation was significantly inhibited,cell migration was decreased,and apoptosis was significantly increased,showing time-dependent and dose-dependent effects.3.After cordycepin ACTS on glioma U87 cells,the expression of KDR mRNA is significantly reduced,which is consistent with the effect of cordycepin on U87 cells,indicating that cordycepin exerts tumor suppressive effect by regulating the expression of KDR. |
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