NF-κB在抗人DR5抗体诱导白血病细胞系凋亡中的作用

Effect of NF-κB on apoptosis of leukemia cell line induced by anti-human DR monoclonal antibody

观察NF-κB在抗人DR5抗体mDRA-6诱导白血病细胞凋亡中的激活,探讨NF-κB在抗人DR5抗体诱导白血病细胞凋亡中的作用。提取细胞蛋白,间接ELISA法检测NF-κB的激活;预先应用NF-κB抑制剂孵育细胞1h,MTT及流式细胞术检测mDRA-6对白血病Jurkat和U937细胞系的凋亡率。结果:mDRA-6作用Jurkat和U937细胞15min,激活形式NF-κB(NF-κBP65)明显增高;10mg/L的mDRA-6作用Jurkat和U937细胞4h,流式细胞术检测细胞凋亡率分别为56.63%和34.14%,而预先用15μmol/L的NF-κB抑制剂处理细胞1h,mDRA-6所致的Jurkat和U937细胞凋亡率分别增加至75.51%和58.01%。mDRA-6诱导白血病Jurkat和U937细胞凋亡过程中激活NF-κB,激活的NF-κB抑制mDRA-6的肿瘤细胞凋亡诱导作用。

To observe the activation of nuclear factor κB （NF-κB）and to investigate the effect of NF-κB on the apoptosis of Jurkat and U937 cells induced by anti-human DR5 monoclonal antibody （mDRA-6）,total proteins of Jurkat and U937 cells was extracted ,and the activated NF-κB of Jurkat and U937 cells was detected with indirect ELISA assay. The apoptotic rate of Jurkat and U937 cells inducing by mDRA-6 were detected by MTT and flow cytometry. It was demonstrated that the level of the activated NF-κB in Jurkat and U937 cells treated with mDRA-6 for 15 minutes was significantly increased,As demonstrated by flow cytometry,the apoptotic rates of Jurkat and U937 cells acted by 10 mg/L of mDRA-6 for 4 hours were 56.63% and 34.14%respectively and the pre-treatment of these two kind of cells with 15 μmol/L of mDRA-6 inhibitor for one hour rendered the apoptotic rates of the affected cells to increase up to 76.51% and 58.01% respectively. It is concluded that anti-human DR5 monoclonal antibody can induce the apoptosis of the leukemic Jurkat and U937 cells through the expression of activated NK-κB and its inhibitor can block this effect.