Activation of MAD 2 checkprotein and persistence of cyclin B1/CDC 2 activity associate with paclitaxel-induced apoptosis in human nasopharyngeal carcinoma cells.
Paclitaxel (Taxol) is a microtubule-interfering agent that induced persistent and transient G2/M arrest before apoptosis in human nasopharyngeal carcinoma (NPC) cells at high and low concentrations, respectively. In this study, we intended to explore the underlying molecular events and found that cellular cyclin B1/CDC 2 kinase activity was increased and persisted for >6 h upon paclitaxel treatment both at high and low concentrations. Furthermore, activation of MAD 2 checkprotein could account for the loss of cyclin B1 ubiquitination and the persistence of cyclin B1/CDC 2 activation in the cases. To investigate the involvement of cyclin B1 and MAD 2 activation in paclitaxel-induced apoptosis, we introduced affinity-purified anti-cyclin B1 and MAD 2 antibodies into NPC cells by electroporation before the further paclitaxel treatment. The antibodies against cyclin B1 and MAD 2 indeed attenuated paclitaxel-induced cytotoxicity and DNA fragmentation. Our study suggests that activation of cyclin B1/CDC 2 and MAD 2 were the M-phase events required for paclitaxel-induced apoptosis in NPC cells. The dys-regulated cyclin B1/CDC 2 activation could enhance the prometaphase progression, but activation of MAD 2 rendered cells inable to exit from the metaphase. Under this circumstance, cells were probably going to "mitotic catastrophe" and ultimately, destined to apoptosis.
Huang
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TS
Cancer Research Division, National Health Research Institutes, Taipei, Taiwan, Republic of China. tshuang@nhri.org.tw
Shu
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CH
Chao
Y
Y
Chen
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Chen
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D000906:Antibodies / Q000276:immunology; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis; D015153:Blotting, Western; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002352:Carrier Proteins; D002453:Cell Cycle / Q000502:physiology; D018797:Cell Cycle Proteins / Q000378:metabolism*; D019926:Cyclin B / Q000378:metabolism*; D056744:Cyclin B1; D053938:DNA Fragmentation; D018274:Electroporation; D005656:Fungal Proteins / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D009303:Nasopharyngeal Neoplasms / Q000378:metabolism / Q000473:pathology*; D009687:Nuclear Proteins; D017239:Paclitaxel / Q000494:pharmacology*; D011233:Precipitin Tests; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000972:Antineoplastic Agents, Phytogenic; C534699:CCNB1 protein, human; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D019926:Cyclin B; D056744:Cyclin B1; D005656:Fungal Proteins; D009687:Nuclear Proteins; D016203:CDC2 Protein Kinase; D017239:Paclitaxel