九节龙皂苷-ⅰ对脑胶质瘤细胞侵袭迁移的作用及机制分析-effect and mechanism analysis of sarsasaponin - ⅰ on invasion and migration of glioma cells.docxVIP

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九节龙皂苷-ⅰ对脑胶质瘤细胞侵袭迁移的作用及机制分析-effect and mechanism analysis of sarsasaponin - ⅰ on invasion and migration of glioma cells.docx

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九节龙皂苷-ⅰ对脑胶质瘤细胞侵袭迁移的作用及机制分析-effect and mechanism analysis of sarsasaponin - ⅰ on invasion and migration of glioma cells

Effects and mechanism of Ardipusilloside-Ⅰon cell invasion andmigration ability of glioma cellsAbstractObjective: To investigate the the influence of Ardipusilloside-I, a triterpenoid saponin isolated from Ardisia pusilla A. DC on invasion and migration of glioma cell line U87 cells and evaluate its mechanism. Methods : U87 cells were cultured without bacteria in vitro, the effect of different concentrationsof Ardipusilloside-I（0、2、4、8、16、32 μM） on U87 cells viability weremeasured by 3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay , then observed the effect on different time（24、48、72h）.The model of transwell chamber which with 50mg?L-1 matrigel-coated for imitatingthe extracellular matrix in vivo was used to investigate the effect of invasion capabilities of U87 cells with different concentrations of Ardipusilloside-I（1、 3、5 μM）after 24h. The migration capabilities of U87 cells were assessed usingwound healing assay after dealing with different concentrations of Ardipusilloside-I（1、3、5 μM）and different time（0、6、24h）in 6-well plates. Matrix-metalloproteinase (MMP) was determined after using different concentrations of Ardipusilloside-I （1、3、5 μM）on U87 cells by gelatin zymography on 8% polyacrylamide gel containing 1g?L-1 gelatin. Throughcorresponding the antibody and antigen, detected the positive reaction of MMPs in U87 cells with antibody at different concentrations Ardipusilloside-I（1、3、5 μM）in immunochemical assay. Results: MTT assay showedArdipusilloside-I inhibited the proliferation of glioma U87 cells, and the tendency in dose-dependent and time-dependent manner, the IC50 valuescalculated from the dose effect curve were 8.02 ± 0.16 μM、6.63 ± 0.10 μM、5.41 ± 0.05 μM for Ardipusilloside-I at 24h, 48h and 72h incubation respectively. Then choose the effective time of Ardipusilloside-I on cells was24h and the drug concentrations were 1、3、5 μM from this assay as standard forother experiments. Compared with control group,