The Study On The Mechanism Of Apoptosis Of Glioma U87 Cells Regulated By The Venom Of Spider Macrothele Raven

Purpose:Gliomas is a tumor occurred in neuroectodermal.It is a serious threat to human health.So far there is no effective treatment to the gliomas.From the various types of gliomas,the most serious is astrocytoma,U87 is one of the glioma cells.Reye Macrothele is a new species of spider which was found recently.In recent years,a large number of experimental studies have shown the venom of spider Macrothele raven has anti-tumor effect,and the proliferation of gastric cancer cells,esophageal cancer cells and cancer cells were inhibited by the venom of spider Macrothele raven.But the mechanism is not fully elucidated.In this experiment,we explore the molecular targeting mechanism of apoptosis of U87 cells targeted by the venom of spider Macrothele raven,and the components with anti-tumor activity were separated and purificated.Lay the foundation for research and development of new anti-cancer drugs.Methods: In this experiment,the model is U87 cells.The U87 cells were treated by the venom of spider Macrothele raven,then we detected the anti-tumor effect by MTS.It will respectively from the cellular level,molecular level to study the venom of spider Macrothele raven regulation on U87 cells.Cellular Level: we can observe the U87 morphological changes of cell apoptosis by inverted microscop and fluorescence staining.The apoptosis of U87 cells was detected by flow cytometry.Molecular Level: The expression of P-ERK?P-JNK?P-p38 detected by Western-blot.Then ERK inhibitor,JNK / SAPK and p38 signal transduction pathway inhibitor were added in U87 cells,and we detected the express of associated protein by Western-blot.The venom of spider Macrothele raven need to be separated and purifed.The first,the venom of spider Macrothele raven was separated by fast protein liquid chromatography to get 1~9 component,the 6 had anti-tumor effect detected by MTS assay,Each component was concentrated and freeze-dried,enrichment.then the component A6 was separated continuely.After all,Detecting the inhibition of various protein components to U87 cells in vitro by MTS assay,the component 6-5 had the strong suppression to U87 cells.Results: The venom of spider Macrothele raven affected U87 cells,inducing apoptosis in U87,cell morphology changes and becomes irregular as pleomorphic.The expression of P-p38 and P-JNK protein increased in U87 cells effected by the venom of spider Macrothele raven.but the P-ERK was opposite.And then ERK inhibitor,JNK / SAPK and p38 signaL transduction pathway inhibitor were added in U87 cells,the result had shown the expression of p-ERK protein increased,and the expression of P-p38 and P-JNK protein was opposite.Reye Macrothele toxin was separated to get 1~9 component,the 6 had anti-tumor effect detected by MTS assay,then the component A6 was separated.After all,detecting the inhibition of various protein components to U87 cells in vitro by MTS assay,the component 6-5 had the strong suppression to U87 cells,the IC50 was 10ug/mL.Conclusion: The venom of spider Macrothele raven can inhibit the proliferation of U87 cells,and it can induce apoptosis by regulating MAPK,JNK / SAPK-MAPK,and p38 pathways.It became the theoretical basis for the development of glioma targeting modulating drugs availabLe.