Tetrandrine Inhibits Proliferation And Motility Of Human Pleomorphic Brain Glioma Cells Through NF-κB Signaling Pathway

Objective:To expound the effects of tetrandrine on the proliferation and movement of human polymorphic glioma GBM8401 cells,and analyze the molecular mechanism of tetrandrine on GBM8401 cells.Methods:The GBM8401 cells stimulated by 5μmol/L or 25μmol/L tetrandrine were used as the experimental group of low or high concentration tetrandrine respectively(5μmol/L TET group,25μmol/L TET group).Adding 10nmol/L Gefitinib(EGFR inhibitor)to each experimental group,as the corresponding EGFR inhibitor group(5μmol/L TET+ EGFR inhibitor group,25μmol/L TET+EGFR inhibitor group).The GBM8401 cells without any stimulation were the control group.The Optical density value(OD value)of GBM8401 cell proliferation in each group was detected by CCK-8.Then the cell wound scratch assay and Transwell cell migration assay were used to detect the change of GBM8401 cell scratch area closure rate and migration number in each group.The content of N-cadherin,L-selectin and IL-8 protein in GBM8401 cells was detected by ELISA.Western Blot was used to detect the expression of N-cadherin,L-selectin,EGFR,p-EGFR,NF-κB and Phosphorylation-NF-κB(p-NF-κB)protein in GBM8401 cells of each group.Results:(1)Compared with the control group,the OD value of GBM8401 cells in the 5 μmol/L or 25 μmol/L experimental group of tetrandrine was significantly decreased,there were all significantly different(P<0.01).Compared with the experimental group of tetrandrine at 5μmol/L or 25μmol/L,OD value of GBM8401 cells was significantly increased in the corresponding 5μmol/L TET+ EGFR inhibitor group,25μmol/L TET+EGFR inhibitor group respectively,there were all significantly different(P<0.01).(2)Compared with the control group,the scratch area closure rate and cell migration number of GBM8401 cells in the experimental group of 5μmol/L or 25 μmol/L tetrandrine were significantly decreased,and there were all significantly different(P<0.01).Compared with the experimental group of tetrandrine at 5μmol/L or 25 μmol/L,the scratch area closure rate and cell migration number of GBM8401 cells were significantly increased in the corresponding 5μmol/L TET+ EGFR inhibitor group,25μmol/L TET+EGFR inhibitor group respectively,there were all significantly different(P<0.01).The GBM8401 cell scratch area closure rate and cell migration rate in each experimental group decreased as the tetrandrine concentration of 5μmol/L or 25 μmol/L increased.(3)Compared with the control group,the expression of N-cadherin,L-selectin or IL-8 protein in 5μmol/L or 25 μmol/L tetrandrine GBM8401 cells of experimental group was significantly decreased(P<0.01).The expression of EGFR,p-EGFR,NF-κB and p-NF-κB in each experimental group were significantly decreased,there were all significantly different(P<0.01).Compared with 5μmol/L or 25 μmol/L tetrandrine experimental groups,respectively,the GBM8401 cells expression of N-cadherin and L-selectin protein significantly increased in the corresponding 5μmol/L TET+ EGFR inhibitor group,25μmol/L TET+EGFR inhibitor group,there were all significantly different(P<0.01).The GBM8401 cells expression of EGFR,p-EGFR,NF-κB,and p-NF-κB protein in corresponding 5μmol/L TET+ EGFR inhibitor group,25μmol/L TET+EGFR inhibitor group was significantly increased,there were all significantly different(P<0.01).Conclusion:(1)Tetrandrine downregulated the OD value of GBM8401 cell proliferation.(2)Tetrandrine significantly inhibited the scratch area closure rate and cell migration number of GBM8401 cells.The changes of these cell movements decreased as the tetrandrine concentration of 5μmol/L or 25 μmol/L increased.(3)Tetrandrine inhibited the expression of EGFR,p-EGFR,NF-κB and p-NF-κB protein in GBM8401 cells.