吉西他滨对9LacZ大鼠脑胶质瘤细胞的生长增殖与凋亡的影响

Effects of Gemcitabine on the proliferation and apoptosis of 9LacZ rat glioma cells

目的研究吉西他滨(Gemcitabine,GEM)对9LacZ大鼠胶质瘤细胞生长增殖与凋亡的影响。方法通过MTT实验检测不同浓度(0.001、0.01、0.1、1、10、100、1000、10000μM)GEM对9LacZ胶质瘤细胞生长的影响。采用DAPI细胞核染色和AnnexinV-FITC/PI染色法检测GEM对9LacZ胶质瘤细胞凋亡的作用。通过Western blot实验检测GEM对9LacZ胶质瘤细胞中caspase-3、Bax和Bcl-2蛋白的调节作用。结果MTT实验表明GEM处理8、12 h,各给药组对9LacZ胶质瘤细胞增殖的抑制作用与对照组相比,差异没有统计学意义(P>0.05);GEM处理24 h,0.001、0.01、0.1μM浓度组对9LacZ胶质瘤细胞增殖的抑制作用与对照组相比,差异无统计学意义(P>0.05);1、10、100、1000、10000μM浓度组对9LacZ胶质瘤细胞增殖的抑制作用与对照组相比,差异有统计学意义(P<0.01)。DAPI细胞核染色实验表明GEM处理24 h,1、10μM干预组细胞核缩小呈不规则状、染色质浓缩发亮。AnnexinV-FITC/PI染色法检测表明:GEM处理24 h,细胞凋亡率1、10μM干预组与对照组相比,差异有统计学意义(P<0.05或P<0.01)。Western blot实验表明:GEM处理24 h,1和10μM浓度组细胞caspase-3蛋白表达明显降低,与对照组相比差异有统计学意义(P<0.01);1和10μM浓度组细胞Bax/Bcl-2的相对蛋白表达率升高较明显,与对照组相比差异有统计学意义(P<0.05,P<0.01)。结论GEM对9LacZ胶质瘤细胞的生长增殖具有抑制作用;GEM可能是通过升高凋亡调控蛋白Bax/Bcl-2的相对蛋白表达率、降低caspase-3的蛋白水平,从而促进9LacZ细胞的凋亡。

Objective To investigate the effects of gemcitabine(GEM)on the proliferation and apoptosis of 9LacZ rat glioma cells.Methods MTT assay was used to measure the growth inhibition of GEM on 9LacZ rat glioma cells at different concentrations(0.001、0.01、0.1、1、10、100、1000、10000μM).DAPI nucleus staining and AnnexinV-FITC/PI staining were used to measure cell apoptosis.The proteins expression of caspase-3、Bax and Bcl-2 were detected by western blot.Results After 8 and 12 hours treatment of GEM,the results of MTT assay showed that different dosage of GEM h ad no effect on the growth of 9LacZ glioma cells compared with control group(P>0.05);After 24 hours treatment of GEM,When compared with control group,the viability of 9LacZ glioma cells showed no significant changes in 0.001,0.01 and 0.1μM GEM-treated groups(P>0.05),but was decreased significantly in 1,10,100,1000 and 10000μM GEM-treated groups(P<0.01).DAPI-staining assay showed that after 24 hours GEM treatment,the nuclear of 1 and 10μM dosage groups narrowed and irregularly shaped,combined with chromatin condenses and brightens.The AnnexinV-FITC/PI staining method showed that:GEM treatment for 24 h,the apoptosis rate was 1,10μM compared with the control group,the difference was statistically significant(P<0.05 or P<0.01).The results of western blot showed that 1 and 10μM GEM-treated groups could significantly decrease the protein levels of caspase-3(P<0.01),and increase the relative protein expression rate of Bax/Bcl-2 compared with control group(P<0.05,P<0.01).Conclusion GEM could inhibit the growth and proliferation of 9LacZ glioma cells by decreasing protein levels of caspase-3 and increasing the relative expression rate of Bax/Bcl-2.