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Activation of MAD 2 checkprotein and persistence of cyclin B1/CDC 2 activity associate with paclitaxel-induced apoptosis in human nasopharyngeal carcinoma cells

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Abstract

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.

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References

  1. Nurse P. Ordering S phase and M phase in the cell cycle. Cell 1994; 79: 547–550.

    PubMed  Google Scholar 

  2. King RW, Jackson PK, Kirschner MW. Mitosis in transition. Cell 1994; 79: 563–571.

    PubMed  Google Scholar 

  3. Fang G, Yu H, KirschnerMW. The checkpoint protein MAD2 and the mitotic regulator CDC20 form a ternary complex with the anaphase-promoting complex to control anaphase initiation. Genes Dev 1998; 12: 1871–1883.

    PubMed  Google Scholar 

  4. Gorbsky GJ, Chen R-H, Murray AW. Microinjection of antibody to Mad2 protein into mammalian cells in mitosis induces premature anaphase. J Cell Biol 1998; 141: 1193–1205.

    PubMed  Google Scholar 

  5. Kallio M, Weinstein J, Daum JR, Burke DJ, Gorbsky GJ. Mammalian p55CDC mediates association of the spindle checkpoint protein Mad2 with the cyclosome/anaphasepromoting complex, and is involved in regulating anaphase onset and late mitotic events. J Cell Biol 1998; 141: 1393–1406.

    PubMed  Google Scholar 

  6. Wassmann K, Benezra R. Mad2 transiently associates with an APC/p55Cdc complex during mitosis. Proc Natl Acad Sci USA 1998; 95: 11193–11198.

    PubMed  Google Scholar 

  7. Sarosy G, Reed E. Taxol dose intensification and its clinical implications. J Natl Med Assoc 1993; 85: 427–431.

    PubMed  Google Scholar 

  8. Forastiere AA, NeubergD, Taylor SG IV, Deconti R, Adams G. Phase II evaluation of Taxol in advanced head and neck cancer: An Eastern Cooperative Oncology Group Trial. Monogr Natl Cancer Inst 1993; 15: 181–184.

    PubMed  Google Scholar 

  9. Schiff PB, Horwitz SB. Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci USA 1980; 77: 1561–1565.

    PubMed  Google Scholar 

  10. Kumar N. Taxol-induced polymerization of purified tubulin. Mechanism of action. J Biol Chem 1981; 256: 10435–10441.

    PubMed  Google Scholar 

  11. Crossin KL, Carney DH. Microtubule stabilization by taxol inhibits initiation of DNA synthesis by thrombin and epidermal growth factor. Cell 1981; 27: 341–350.

    PubMed  Google Scholar 

  12. Jordan MA, Toso RJ, Thrower D, Wilson L. Mechanism of mitotic block and inhibition of cell proliferation by taxol at low concentrations. Proc Natl Acad Sci USA 1993; 90: 9552–9556.

    PubMed  Google Scholar 

  13. Shu C-H, Yang WK, Shih Y-L, Kuo M-L, Huang T-S. Cell cycle G2/M arrest and activation of cyclin-dependent kinases associated with low-dose paclitaxel-induced sub-G1 apoptosis. Apoptosis 1997; 2: 463–470.

    PubMed  Google Scholar 

  14. Shen S-C, Huang T-S, Jee S-H, Kuo M-L. Taxol-induced p34cdc2 kinase activation and apoptosis inhibited by 12-Otetradecanoylphorbol-13-acetate in human breast MCF-7 carcinoma cells. Cell Growth Differ 1998; 9: 23–29.

    PubMed  Google Scholar 

  15. Halicka HD, Seiter K, Feldman EJ, et al. Cell cycle specificity of apoptosis during treatment of leukemias. Apoptosis 1997; 2: 25–39.

    PubMed  Google Scholar 

  16. Shu C-H, Yang WK, Huang T-S. Increased cyclin B1/CDC 2 kinase activity and phosphorylation of Bcl-2 associated with paclitaxel-induced apoptosis in human nasopharyngeal carcinoma cells. Apoptosis 1996; 1: 141–146.

    Google Scholar 

  17. Huang T-S, Kuo M-L, Shew J-Y, Chou Y-W, Yang WK. Distinct p53-mediated G1/S checkpoint responses in two NIH3T3 subclone cells following treatment with DNAdamaging agents. Oncogene 1996; 13: 625–632.

    PubMed  Google Scholar 

  18. Huang T-S, Duyster J, Wang JYJ. Biological response to phorbol ester determined by alternative G1 pathway. Proc Natl Acad Sci USA 1995; 92: 4793–4797.

    PubMed  Google Scholar 

  19. Huang T-S, Shu C-H, Yang WK, Whang-Peng J. Activation of CDC 25 phosphatase and CDC 2 kinase involved in GL331-induced apoptosis. Cancer Res 1997; 57: 2974–2978.

    PubMed  Google Scholar 

  20. Huang T-S, Yang WK, Whang-Peng J. GL331-induced disruption of cyclin B1/CDC 2 complex and inhibition of CDC 2 kinase activity. Apoptosis 1996; 1: 213–217.

    Google Scholar 

  21. Shi L, Nishioka WK, Th'ng J, Bradbury EM, Lichfield DW, GreenbergAH. Premature p34cdc2 activation required for apoptosis. Science 1994; 263: 1143–1145.

    PubMed  Google Scholar 

  22. Fotedar R, Flatt J, Gupta S, et al. Activation-induced T-cell death is cell cycle dependent and regulated by cyclin B. Mol Cell Biol 1995; 15: 932–942.

    PubMed  Google Scholar 

  23. Shimizu T, O'connor PM, KohnKW, Pommier Y. Unsheduled activation of Cyclin B1/Cdc 2 kinase in human promyelocytic leukemia cell line HL60 cells undergoing apoptosis induced by DNA damage.Cancer Res 1995; 55: 228–231.

    PubMed  Google Scholar 

  24. Zhou B-B, Li H, Yuan J, Kirschner MW. Caspase-dependent activation of cyclin-dependent kinases during Fas-induced apoptosis in Jurkat cells. Proc Natl Acad Sci USA 1998; 95: 6785–6790.

    PubMed  Google Scholar 

  25. Peter M, Nagagawa J, Doree M, Labbe JC, Nigg EA. In vitro disassembly of the nuclear lamina and M phase-specific phosphorylation of lamins by cdc2 kinase. Cell 1990; 61: 591–602.

    PubMed  Google Scholar 

  26. Ward GE, KirschnerMW. Identification of cell cycle-regulated phosphorylation sites on nuclear lamin C. Cell 1990; 61: 561–577.

    PubMed  Google Scholar 

  27. Chou Y-H, Bischoff JR, Beach D, Goldman RD. Intermediate filament reorganization during mitosis is mediated by p34cdc2 phosphorylation of vimentin. Cell 1990; 62: 1063–1071.

    PubMed  Google Scholar 

  28. Cai S-Y, Babbitt RW, Marchesi VT. A mutant deubiquitinating enzyme (Ubp-M) associates with mitotic chromosomes and blocks cell division. Proc Natl Acad Sci USA 1999; 96: 2828–2833.

    PubMed  Google Scholar 

  29. Walker PR, Sikorska M. New aspects of the mechanism of DNA fragmentation in apoptosis. Biochem Cell Biol 1997; 75: 287–299.

    PubMed  Google Scholar 

  30. Kratzmeier M, Albig W, Meergans T, Doenecke D. Changes in the protein pattern of H1 histones associated with apoptotic DNA fragmentation. Biochem J 1999; 337: 319–327.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Cooperative Laboratory, Cancer Research Division, National Health Research Institutes, Taipei, Taiwan, Republic of China

    Tze-Sing Huang

  2. Department of Otolaryngology, Veterans General Hospital, Taipei, Taiwan, Republic of China

    Chih-Hung Shu

  3. Cancer Center, Veterans General Hospital, Taipei, Taiwan, Republic of China

    Yee Chao

  4. Cooperative Laboratory, Cancer Research Division, National Health Research Institutes, Taipei, Taiwan, Republic of China

    San-Na Chen & Li-Li Chen

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  1. Tze-Sing Huang
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  2. Chih-Hung Shu
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Huang, TS., Shu, CH., Chao, Y. et al. Activation of MAD 2 checkprotein and persistence of cyclin B1/CDC 2 activity associate with paclitaxel-induced apoptosis in human nasopharyngeal carcinoma cells. Apoptosis 5, 235–241 (2000). https://doi.org/10.1023/A:1009652412399

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