Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지

Apoptotic Cell Death in TrkA-overexpressing Cells: Kinetic Regulation of ERK Phosphorylation and Caspase-7 Activation

  • Jung, Eun Joo (Department of Biochemistry and RINS, MRCND and Institute of Health Sciences, Gyeongsang National University School of Medicine) ;
  • Kim, Deok Ryong (Department of Biochemistry and RINS, MRCND and Institute of Health Sciences, Gyeongsang National University School of Medicine)
  • Received : 2007.10.17
  • Accepted : 2008.01.23
  • Published : 2008.07.31
⟨ Previous Next ⟩

Abstract

The TrkA tyrosine kinase is activated by autophosphorylation in response to NGF, and plays an important role in cell survival, differentiation, and apoptosis. To investigate its role in cell fate determination, we produced stable TrkA-inducible SK-N-MC and U2OS cell lines using the Tet-On system. Interestingly, TrkA overexpression induced substantial cell death even in the absence of NGF, by stimulating ERK phosphorylation and caspase-7 activation leading to PARP cleavage. TrkA-mediated cell death was shown by the annexin-V binding assay to be, at least in part, apoptotic in both SK-N-MC and U2OS cells. Furthermore, the truncated form (p18) of Bax accumulated in the TrkA-induced cells, suggesting that TrkA induces mitochondria-mediated apoptosis. NGF treatment augmented the cell death induced by TrkA overexpression. This TrkA-induced cell death was blocked by the tyrosine kinase inhibitors, K-252a and GW441756. Moreover, TrkA overexpression inhibited long-term proliferation of both the neuronal SK-N-MC cells and the non-neuronal U2OS cells, suggesting a potential role of TrkA as a tumor suppressor.

Keywords

Acknowledgement

Supported by : Korea Research Foundation, Korea Science and Engineering Foundation

References

  1. Cartron, P.F., Oliver, L., Juin, P., Meflah, K., and Vallette, F.M. (2004). The p18 truncated form of Bax behaves like a Bcl-2 homology domain 3-only protein. J. Biol. Chem. 279, 11503- 11512 https://doi.org/10.1074/jbc.M311922200
  2. Chou, T.T., Trojanowski, J.Q., and Lee, V.M. (2000). A novel apoptotic pathway induced by nerve growth factor-mediated TrkA activation in medulloblastoma. J. Biol. Chem. 275, 565-570 https://doi.org/10.1074/jbc.275.1.565
  3. Costantini, C., Scrable, H., and Puglielli, L. (2006). An aging pathway controls the TrkA to p75NTR receptor switch and amyloid beta-peptide generation. EMBO J.25, 1997-2006 https://doi.org/10.1038/sj.emboj.7601062
  4. Cowley, S., Paterson, H., Kemp, P., and Marshall, C.J. (1994). Activation of MAP kinase kinase is necessary and sufficient for PC12 differentiation and for transformation of NIH 3T3 cells. Cell 77, 841-852 https://doi.org/10.1016/0092-8674(94)90133-3
  5. Friedman, W.J., and Greene, L.A. (1999). Neurotrophin signaling via Trks and p75. Exp. Cell. Res.253, 131-142 https://doi.org/10.1006/excr.1999.4705
  6. Jung, E.J., and Flemington, E.K. (2001). Transfection-mediated cell synchronization: acceleration of G1-S phase transition by gamma irradiation. BioTechniques 31, 1026, 1028, 1031-1024
  7. Klein, R., Jing, S.Q., Nanduri, V., O'Rourke, E., and Barbacid, M. (1991). The trk proto-oncogene encodes a receptor for nerve growth factor. Cell 65, 189-197 https://doi.org/10.1016/0092-8674(91)90419-Y
  8. Kurz, E.U., and Lees-Miller, S.P. (2004). DNA damage-induced activation of ATM and ATM-dependent signaling pathways. DNA Repair 3, 889-900 https://doi.org/10.1016/j.dnarep.2004.03.029
  9. Kurz, E.U., Douglas, P., and Lees-Miller, S.P. (2004). Doxorubicin activates ATM-dependent phosphorylation of multiple downstream targets in part through the generation of reactive oxygen species. J. Biol. Chem. 279, 53272-53281 https://doi.org/10.1074/jbc.M406879200
  10. Marshall, C.J. (1995). Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell 80, 179-185 https://doi.org/10.1016/0092-8674(95)90401-8
  11. Muragaki, Y., Chou, T.T., Kaplan, D.R., Trojanowski, J.Q., and Lee, V.M. (1997). Nerve growth factor induces apoptosis in human medulloblastoma cell lines that express TrkA receptors. J. Neurosci. 17, 530-542 https://doi.org/10.1523/JNEUROSCI.17-02-00530.1997
  12. Reichardt, L.F. (2006). Neurotrophin-regulated signalling pathways. Phil. Trans. R. Soc. B. 361, 1545-1564 https://doi.org/10.1098/rstb.2006.1894
  13. Rong, J., McGuire, J.R., Fang, Z.H., Sheng, G., Shin, J.Y., Li, S.H., and Li, X.J. (2006). Regulation of intracellular trafficking of huntingtin- associated protein-1 is critical for TrkA protein levels and neurite outgrowth. J. Neurosci. 26, 6019-6030 https://doi.org/10.1523/JNEUROSCI.1251-06.2006
  14. Schramm, A., Schulte, J.H., Astrahantseff, K., Apostolov, O., Limpt, V., Sieverts, H., Kuhfittig-Kulle, S., Pfeiffer, P., Versteeg, R., and Eggert, A. (2005). Biological effects of TrkA and TrkB receptor signaling in neuroblastoma. Cancer Lett. 228, 143-153 https://doi.org/10.1016/j.canlet.2005.02.051
  15. Shikata, A., Shikata, T., Sotozono, Y., Hosoi, H., Matsumura, T., Sugimoto, T., and Sawada, T. (2000). Neuronal differentiation in human neuroblastoma cells by nerve growth factor following TrkA up-regulation by interferon-gamma. Med. Pediatr. Oncol. 34, 394-401 https://doi.org/10.1002/(SICI)1096-911X(200006)34:6<394::AID-MPO3>3.0.CO;2-5
  16. Shim, H.Y., Park, J.H., Paik, H.D., Nah, S.Y., Kim, D.S., and Han, Y.S. (2007). Acacetin-induced apoptosis of human breast cancer MCF-7 cells involves caspase cascade, mitochondriamediated death signaling and SAPK/JNK1/2-c-Jun activation. Mol. Cells 24, 95-104
  17. Stephens, R.M., Loeb, D.M., Copeland, T.D., Pawson, T., Greene, L.A., and Kaplan, D.R. (1994). Trk receptors use redundant signal transduction pathways involving SHC and PLC-gamma 1 to mediate NGF responses. Neuron 12, 691-705 https://doi.org/10.1016/0896-6273(94)90223-2
  18. Tacconelli, A., Farina, A.R., Cappabianca, L., Desantis, G., Tessitore, A., Vetuschi, A., Sferra, R., Rucci, N., Argenti, B., Screpanti, I., et al. (2004). TrkA alternative splicing: a regulated tumor-promoting switch in human neuroblastoma. Cancer Cell 6, 347-360 https://doi.org/10.1016/j.ccr.2004.09.011
  19. Tang, D., Wu, D., Hirao, A., Lahti, J.M., Liu, L., Mazza, B., Kidd, V.J., Mak, T.W., and Ingram, A.J. (2002). ERK activation mediates cell cycle arrest and apoptosis after DNA damage independently of p53. J. Biol. Chem.277, 12710-12717 https://doi.org/10.1074/jbc.M111598200
  20. Toyota, H., Yanase, N., Yoshimoto, T., Moriyama, M., Sudo, T., and Mizuguchi, J. (2003). Calpain-induced Bax-cleavage product is a more potent inducer of apoptotic cell death than wild-type Bax. Cancer Lett.189, 221-230 https://doi.org/10.1016/S0304-3835(02)00552-9
  21. Urra, S., Escudero, C.A., Ramos, P., Lisbona, F., Allende, E., Covarrubias, P., Parraguez, J.I., Zampieri, N., Chao, M.V., Annaert, W., et al. (2007). TrkA receptor activation by nerve growth factor induces shedding of the p75 neurotrophin receptor followed by endosomal gamma-secretase-mediated release of the p75 intracellular domain. J. Biol. Chem.282, 7606-7615 https://doi.org/10.1074/jbc.M610458200
  22. Woessmann, W., Chen, X., and Borkhardt, A. (2002). Rasmediated activation of ERK by cisplatin induces cell death independently of p53 in osteosarcoma and neuroblastoma cell lines. Cancer Chemother. Pharmacol.50, 397-404 https://doi.org/10.1007/s00280-002-0502-y
  23. Wood, D.E., Thomas, A., Devi, L.A., Berman, Y., Beavis, R.C., Reed, J.C., and Newcomb, E.W. (1998). Bax cleavage is mediated by calpain during drug-induced apoptosis. Oncogene 17, 1069-1078 https://doi.org/10.1038/sj.onc.1202034
  24. Yan, C., Liang, Y., Nylander, K.D., and Schor, N.F. (2002). TrkA as a life and death receptor: receptor dose as a mediator of function. Cancer Res.62, 4867-4875