The Mechanism Of Nimesulide Induces Glioma U87Cell Lines To Apoptosis

Objective: To investigate the effect of Nimesulide affecting proliferationand apoptosis of glioma U87cell lines by different methods in vitro, and toexplore the mechanism of its effects.Methods: U87glioblatoma cell lines were cultured for18hours, aftergrouping with differentcon centrations of Nimesulide:0.05,0.1,0.2,0.4,0.8mmol/L; grouping with different time:12,24,48hours; and then caculating theinhibination rate of U87glioblastoma cell lines by methylthiazolyl tetrazolium(MTT) method;using acridine orange (AO)/ethidium bromide (EB) stainingand flow cutometry methods to monitor the change of U87glioblastoma celllines; observing morphological changes of apoptosis rate of U87glioblastomacell lines with transmission electron microscopy and AnnexinV-FITC/PI assay;By Westem blotting observing the expression of Bcl-2, Bax and Cox-2protein inU87human glioblastoma cell lines.Results:(1).To analysize U87cell lines which were respectively culturedwith nimesulide solutions containing different concentrations for12,24,48hoursby MTT colorimetric analysis.The growth of U87cells is obviously inhibitedand positively be correlated with different drug-doses and active times.(2).Through acridine orange (AO)/ethidium bromide (EB) staining andtransmissive fluorescence microscopy, we observed that nimesulide made U87glioma cells a significant apoptotic morphological changes;Apoptosis wasoberseved by AnnexinV-FITC/PI assay,which showed that nimesulide madeU87cells apoptosis mainly in the late period, and with increasing concentrationof nimesulide, apoptosis rate was also significantly elevated.(3). ThroughWestern blot gel electrophoresis we found nimesulide not only could dose-dependently reduce the expression of Bcl-2, Cox-2proteins, while increaseBax protein in U87cells which were developed with different nimesulidesolutions containing0.2,0.4,0.8mmol/L for24hours.Conclusion:(1). Nimesulide could effectively inhibit the growth of gliomaU87cell lines in vitro and inhibition rate was dependent with the effective timeand dose of drug.(2). Nimesulide could promote U87cells apoptosis mainly inthe late period, and the cell apoptosis rate is in dose-depentent manner.(3).Nimesulide could significantly inhibit the proliferation and induce apoptosis ofglioma U87cell lines,which its mechanism of apoptosis probably is throughinhibiting the expression of Cox-2and down-regulating Bcl-2andsimultaneously up-regulating Bax protein to achieve its effect of anti-tumor.