| Caspase-dependent molecular mechanisms of anti-human DR5 monoclonal antibody mDRA-6 inducing apoptosis of human leukemia Jurkat cells. | |
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MedLine Citation:
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PMID: 19550122 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND AND OBJECTIVE: Both tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and some monoclonal agonistic antibodies against TRAIL receptors have antitumor activity. We have previously prepared a novel monoclonal agonistic antibody against human death receptor 5 (DR5) and designated it as mDRA-6. This study was to explore the Caspase-dependent molecular mechanisms of mDRA-6 inducing apoptosis of human leukemia Jurkat cells. METHODS: After exposure to different doses of mDRA-6, DNA fragmentation of Jurkat cells was detected by agarose gel electrophoresis, cell proliferation was detected by MTT assay, and cell apoptosis was detected by flow cytometry after Annexin V-FITC/PI double staining. Jurkat cells were further treated with the inhibitors for Caspase-10, -9, -8 and -3. The active cleavage products of Caspase-10, -9, -8, -3 and poly ADP-ribose polymerase (PARP), BH3 interacting domain death agonist (Bid), truncated Bid (tBid) and cytochrome c (Cyto c), were analyzed by western blot. RESULTS: After mDRA-6 treatment, DNA fragmentation was detected in Jurkat cells. mDRA-6 inhibited cell proliferation in a dose-dependent manner. When treated with 2.0 microg/mL mDRA-6, the apoptosis rates of Jurkat cells were 16.2% at 0.25 h, 28.3% at 0.5 h, 69.2% at 1 h and 78.2% at 2 h. Interestingly, the mDRA-6-induced apoptosis was repressed by 77.9% by Caspase-8 inhibitor ZIF, 54.2% by Caspase-3 inhibitor ZDF, and 8.7% by Caspase-9 inhibitor ZLF, but was not repressed by Caspase-10 inhibitor ZAF. After mDRA-6 exposure, the proenzymes of Caspase-8, -9 and -3 were reduced and their active cleavage products were increased along with the increase of exposure time, the cleavage products of PARP were also increased, Bid was degraded to tBid, and an abundance of Cyto c was released from mitochondria, but the proenzyme of Caspase-10 showed no change and no cleavage products of Caspase-10 were detectable. CONCLUSION: mDRA-6 can induce apoptosis of Jurkat cells via the Caspase-dependent and mitochondrial pathways. |
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Authors:
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Yao-Wu Du; Guang-Chao Liu; Jing Wang; Yue-Ping Zhao; Shu-Lian Li; Ju-Gao Chen; Qi Jiang; Jing Cai; Yuan-Fang Ma |
Publication Detail:
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Type: Journal Article Date: 2009-02-27 |
Journal Detail:
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Title: Ai zheng = Aizheng = Chinese journal of cancer Volume: 28 ISSN: 1000-467X ISO Abbreviation: Ai Zheng Publication Date: 2009 Feb |
Date Detail:
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Created Date: 2009-06-26 Completed Date: 2009-10-22 Revised Date: 2010-02-04 |
Medline Journal Info:
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Nlm Unique ID: 9424852 Medline TA: Ai Zheng Country: United States |
Other Details:
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Languages: eng Pagination: 112-6 Citation Subset: IM |
Affiliation:
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Institute of Immunology, Medical College, Henan University, Kaifeng, Henan, PR China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Antibodies, Monoclonal
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pharmacology* Apoptosis / drug effects* BH3 Interacting Domain Death Agonist Protein / metabolism Blotting, Western Caspase 10 / antagonists & inhibitors, metabolism Caspase 3 / antagonists & inhibitors, metabolism Caspase 8 / antagonists & inhibitors, metabolism Caspase 9 / antagonists & inhibitors, metabolism Caspases / antagonists & inhibitors, metabolism* Cell Proliferation / drug effects Cytochromes c / metabolism DNA Fragmentation / drug effects Dose-Response Relationship, Drug Electrophoresis, Agar Gel Flow Cytometry Humans Jurkat Cells Leukemia / enzymology, pathology Oligopeptides / pharmacology Poly(ADP-ribose) Polymerases / metabolism Receptors, TNF-Related Apoptosis-Inducing Ligand / immunology* Time Factors |
| Chemical | |
Reg. No./Substance:
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0/Antibodies, Monoclonal; 0/BH3 Interacting Domain Death Agonist Protein; 0/BID protein, human; 0/Oligopeptides; 0/Receptors, TNF-Related Apoptosis-Inducing Ligand; 0/benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; 0/benzyloxycarbonyl-alanyl-glutamyl-valyl-aspartic acid fluoromethyl ketone; 0/benzyloxycarbonyl-isoleucyl-glutamyl-threonyl-aspartic acid fluoromethyl ketone; 0/benzyloxycarbonyl-leucyl-glutamyl-histidyl-aspartic acid fluoromethyl ketone; 9007-43-6/Cytochromes c; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 3.4.22.-/Caspase 10; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 8; EC 3.4.22.-/Caspase 9; EC 3.4.22.-/Caspases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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