The Effect Of Iron Ion Resist Gossypol Acetate To Kill Glioma Cells

Objective:This study is aimed to investigate the effect of Fe³⁺in human glioma microenvironment on the drug sensitivity of GAA,to provide a basis for further clinical translational research.Methods:Bioinformatics was applied to analyze of TFRC protein and gene expression level in glioma and its clinical significance,as well as the changes of Fe³⁺content in glioma microenvironment.Using different concentrations of the GAA（0,5,10,15,20,25μmol/L）to treat with the human brain glioma LN229,SW1783,U87MG,U251MG cells,cell activity was detected by CCK8 method,and calculate the LD50 of GAA.Glioma cells were then treated in the following four groups:control group,Fe³⁺group,GAA group,Fe³⁺+GAA group.CCK8 cytotoxicity assay was used to detect the cell activity.Transwell assay was used to detect the changes of cell migration ability.To verify the role of Fe³⁺in GAA resistance of glioma,Western Blot and quantitative real-time PCR were used to detect the expression of related apoptotic indexes of glioma cells under different treatment conditions.The apoptosis level of glioma cells was analyzed by flow cytometry.Results:The results of clinical data analysis and bioinformatics analysis showed that the m RNA and protein levels of TFRC in glioma were higher than those in normal brain tissue,and the gene expression of TFRC increased with the increase of malignant degree of glioma.The overall survival rate of the patients with high TFRC expression was lower,suggesting that the expression level of TFRC was negatively correlated with the prognosis of the patients.GAA can significantly inhibit the activity of glioma cells,and the LD50 of GAA to LN229,SW1873,U87MG,U251MG was 9.38μmol/L,9.56μmol/L,14.53μmol/L,10.39μmol/L.In human glioma LN229,SW1783,U87MG,U251MG cells,the effect of Fe³⁺on cell proliferation was not significant compared with the control group,while GAA significantly inhibited the proliferation and migration of glioma cells.Compared with GAA group,the vitality and migration of tumor cells were significantly enhanced after Fe³⁺and GAA treatment,and the combination of GAA and DFO also showed a significant killing effect on glioma cells,even in the presence of Fe³⁺,it can also ensure the killing of brain glioma cells.In human glioma LN229 cells,the expression level of apoptosis-related protein Bcl-2 decreased after treated with GAA,while the expression level of Bcl-2 protein increased relatively after the cells were treated with Fe³⁺and GAA at the same time.In LN229 cells and U251MG cells,the results of flow cytometry also showed that the level of apoptosis in the GAA treatment group increased,while that of the Fe³⁺and GAA treatment group decreased.Conclusions:The high expression of TFRC was negatively correlated with the survival of glioma patients,and it suggested the enrichment of Fe³⁺in glioma.Fe³⁺resists the inhabitation of proliferation and migration caused by GAA in glioma.Fe³⁺resists the apoptosis caused by GAA in glioma.In this study,the effects of Fe³⁺on GAA drug sensitivity in glioma were investigated,and the causes of GAA drug resistance in clinical glioma were explained,which provided a theoretical basis and a new direction for glioma treatment.