【摘要】 目的通过引入自噬及凋亡抑制剂观察三氧化二砷(As2O3)作用于骨髓瘤细胞RPMI 8226引起自噬和凋亡之间的关系。方法①MTT法计算细胞生长抑制率。②流式细胞仪检测RPMI 8226细胞的自噬率及凋亡率的变化。③通过Western Blot方法检测不同实验组Beclin-1及Caspase-3表达的变化。结果①各浓度As2O3对人骨髓瘤细胞RPMI 8226都显示出明显的生长抑制作用(P<0.05),且呈时间-剂量依赖性。②与As2O3单独处理组相比,3-MA预处理组,As2O3诱导RPMI 8226细胞凋亡率呈下降趋势,有统计学意义(P<0.05);zVAD-fmk与As2O3联合处理组,凋亡率未见明显变化(P>0.05)。与As2O3单独处理组相比,3-MA预处理组,As2O3诱导RPMI 8226细胞自噬率下降,有统计学意义(P<0.05);zVAD-fmk与As2O3联合处理组As2O3诱导RP-MI8226细胞自噬率亦下降,有统计学意义(P<0.05)。③与As2O3单独加药组比较,3-MA预处理组As2O3诱导RPMI 8226细胞Procaspase-3蛋白表达明显升高,Beclin-1表达下降;zVAD-fmk预处理组亦表现为Procaspase-3蛋白表达明显升高,Beclin-1下降。结论①不同浓度组As2O3作用24、48、72h对RPMI8226细胞的增殖均有明显的抑制作用(P<0.05),且该作用成时间-剂量依赖关系。②As2O3可诱导RPMI 8226细胞发生凋亡及自噬,两者的关系表现为互为前提。抑制任何一方都会导致另一方过程受抑。③As2O3诱导RPMI 8226细胞凋亡过程中可能存在Caspase非依赖型细胞凋亡程序。更多还原

【Abstract】 Objective To observed the relationship of autophagy and apoptosis induced by arsenic rioxide in human multiple myeloma cell RPMI 8226 by joining the autophagy inhibitor and apoptosis inhibitors.Methods① MTT bioassay was used to analyze the growth inhibition rate.②FACS was used to test the autophagy rate and apoptosis rate.③ Using Western blot to test the expression of Beclin 1、Bcl-2 and Caspase-3 in RPMI 8226 of each group.Results:① Different concentration of As2O3 can significantly inhibit the proliferation of RPMI 8226(P<0.05),and the inhibitory effect resulted in dose and time-dependent inhibition.② Compared with the group treatment with As2O3 alone,the apoptpsis rate decreased in RPMI 8226 cells of the group was treated with 3-MA and As2O3(P<0.05);the apoptosis rate of zVAD-fmk and As2O3 combined treatment group did not change(P>0.05);the autophagy rate decreased in RPMI 8226 cells of the group which treatment with 3-MA and As2O3(P<0.05);the autophagy rate of zVAD-fmk and As2O3 combined treatment group decreased too(P<0.05).(data analysis to remove 3-MA and zVAD-fmk separate treatment effects).③ Compared with the group treatment with As2O3 alone,the protein expression of Procaspase-3 was significantly increased,Beclin-1 decreased in RPMI 8226 cells was pretreated with 3-MA;zVAD-fmk pretreatment group also showed the protein expression of Procaspase-3 was significantly increased,Beclin-1 decreased.Conclusion① Different concentrations of As2O3,treated RPMI 8226 for 24,48,72 h the proliferation of RPMI 8226 inhibited significantly(P<0.05),the inhibitory effect was dose and time-dependent.② As2O3 can induce apoptosis and autophagy in RPMI 8226 cells,the relationship between the apoptosis and autophagy is they are both the premise of each other.Inhibition of either party will result in suppression of the other process.③As2O3 induced apoptosis in RPMI 8226 cells may exist another apoptosis pathway which is a caspase-independent process.更多还原