The mitochondrial-dependent molecular mechanisms for inducing apoptosis in Jurkat cells by a novel agonistic anti-human DR5 monoclonal antibody
- VernacularTitle:抗人DR5功能性单克隆抗体诱导Jurkat细胞凋亡的线粒体途径的分子机制
- Author:
Yaowu DU
;
Lihui CHAI
;
Hongying HUANG
;
Huiling BAI
;
Yueping ZHAO
;
Yuanfang MA
- Publication Type:Journal Article
- Keywords:
Death recptor 5;
Monoclonal antibody;
Jurkat cell;
Apoptosis;
Westem blot
- From:
Chinese Journal of Immunology
2010;26(1):3-7
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate mitochondrial-dependent molecular mechanisms of a novel agonistic anti-human death receptor 5 (DR5) monoclonal antibody(mDRA-6) inducing apoptosis in Jurkat cell.Methods:The dose-dependent and time-dependent cell growth suppression of mDRA-6 in Jurkat cells was determined by MTT assay.The measurement of the mitochondrial transmembrane potential(ΔΨm) of Jurkat cells was detected by flow cytometry with JC-1 single staining.Caspase-8,9 as well as Bid,Bax,Bcl-2 and Cyto c of apoptotic Jurkat cells were analyzed by Western blot after mDRA-6 treatment.Results:The mDRA-6 induced cell growth suppression and cytotoxicity in dose-dependent manner and time-dependent manner.After mDRA-6 treatment at 2.0 μg/ml for15 min,30 min,60 min and 120 min,the change in ΔΨm were 20.14 %,19.34 %,21.11% and 30.90% respectively by JC-1 single staining.Western blot revealed that the level of active fragments of Caspase-8,9 and Bid,Bax,Bcl-2 and Cyto c respectively,and the amount of Cyto c was increased in cytosol concomitant with the related attenuation of Cyto c in mitochondria.Conclusion: Apoptotic pathway of Jurkat cells induced by mDRA-6 is initiated upon DR5 ligatian to mDRA-6 and exogenic Caspase-dependent cell apoptotic cascades is activated,and endogenic mitochondrial-dependent cell apoptosis pathway is activated.mDRA-6 may be a useful agent in investigating human leukemia therapy by using TRAIL/DR5.
