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目的探讨鼠抗人DR5单克隆抗体(mDRA-6)对白血病细胞U937凋亡诱导作用及机制。方法 MTT法、Annexin V-FITC/PI双染流式细胞仪检测mDRA-6对U937细胞的生长抑制及凋亡诱导作用,Western blotting检测caspase8、10、3、9及Cytochome C在U937细胞凋亡过程中的表达及激活情况;选用caspase8、10、3抑制剂预处理U937细胞,观察是否抑制mDRA-6对U937细胞的凋亡诱导作用。结果MTT结果显示:10mg/L的mDRA-6作用U937细胞24h,细胞死亡率为61.09%,呈时间、浓度依赖性;流式细胞仪检测显示mDRA-6作用4h,细胞凋亡率为69.03%;Western blotting结果显示caspase10、3、9及Cytochome C均有活性片断表达,而caspase8无明显激活;caspase10和caspase3抑制剂部分抑制mDRA-6的凋亡诱导作用,caspase8抑制剂作用不明显。结论抗人DR5单克隆抗体mDRA-6通过死亡受体和线粒体途径诱导白血病U937细胞凋亡。
Objective To investigate the apoptosis-inducing effect of murine anti-human DR5 monoclonal antibody (mDRA-6) on leukemia cell line U937 and its mechanism. Methods The growth inhibition and apoptosis induction of mDRA-6 on U937 cells were detected by MTT assay and Annexin V-FITC / PI double staining. The apoptosis of U937 cells induced by caspase8,10,3,9 and Cytochome C were detected by Western blotting U937 cells were pretreated with caspase8,10,3 inhibitor to observe whether they could inhibit the apoptosis-inducing effect of mDRA-6 on U937 cells. Results The results of MTT showed that mDRA-6 treated with 10 mg / L of mDRA-6 for 24 h had a cell viability of 61.09% in a time-and dose-dependent manner. Flow cytometry showed that mDRA-6 treated for 4 h had an apoptotic rate of 69.03% ; Western blotting results showed that both caspase10,3,9 and Cytochome C were active fragments expression, and caspase8 no significant activation; caspase10 and caspase3 inhibitors partially inhibited the induction of mDRA-6 apoptosis, caspase8 inhibitor effect was not obvious. Conclusion The anti-human DR5 monoclonal antibody mDRA-6 induces the apoptosis of leukemia U937 cells through death receptor and mitochondrial pathway.