The Effect Of IDH1^R132H Mutation On Proliferation,Migration Of Glioma Cells

Objective:Glioma is the intracranial tumor that originated in glial cells,accounting for more than 80%of malignant tumors in the central nervous system.Because of its high recurrence,disability and fatality rate,glioma has become one of the difficult problems in the world,which seriously threatens human health.Isocitrate dehydrogenase?IDH?gene mutation is common in low grade gliomas or part of secondary glioblastomas,which have better prognosis.IDH mutation has become an important biomarker of molecular classification of glioma.This study intends to explore the biological behavior of IDH1 mutant(IDH1^R132H)and wild type(IDH1_wt)glioma cells,and then established glioma cell lines which stable expressed the IDH1^R132H and IDH1_wtt protein.This may lay the foundation of further study in the clinical characteristics and its molecular mechanism of IDH mutation gliomas,so as to provide the basis for clinical treatment and prognosis of glioma patients.Methods:The IDH1^R132H and IDH1_wt plasmid were transfected into cells by Lipofectamine 3000 transfection agent,and then using Western blot experiment to detect transfection efficiency.The cell proliferation ability was detected by CCK-8 method and clone formation assay.The migration ability was detected by cell scratch test.The stably transfected cell lines of IDH1^R132H132H and IDH1_wt were screened by puromycin,and Western blot method was applied to verify.Results:Results of Western blot experiment showed that IDH plasmid has been successfully transfected into glioma cells.Results of CCK-8,clone formation assay and cell scratch test respectively showed that overexpression of IDH1^R132H group cells have significantly lower proliferation ability and migration ability than that of the control group and overexpression of IDH1_wtt group.After the screening of puromycin,the stably infected cell lines were established successfully,and Western blot experiment confirmed that the IDH protein with FLAG label expression was detected.Conclusions:Glioma cells with IDH1^R132H mutant have lower abilities of proliferation,migration.This implied IDH mutation herald a relatively low-grade malignant glioma cells.