Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Epidermal growth factor and serum activate distinct pathways to inhibit the BH3 only protein BAD in prostate carcinoma LNCaP cells

Abstract

A better understanding of pathways involved in survival of prostate cancer cells is the key to develop effective and target-selective therapies. Presence of serum or epidermal growth factor in the culture medium of LNCaP cells decreases apoptosis induced by the inhibition of phosphatidylinositol 3-kinase with LY294002. However, intracellular pathway(s) involved in this survival signaling are not well defined. Here, we investigated the mechanism(s) involved in serum or epidermal growth factor-mediated inhibition of LY294002-induced death in LNCaP cells. Cell death was assessed by the percentage of cells in sub-G1 phase and caspase 3 activity. Phosphorylation status of BAD, ERK1/2 and RSKs were assessed by Western blot. Specific gene expression knock down of BAD, BAX, RSK1 and RSK2 were performed using siRNA transfections. Our results demonstrate that cell death induced by LY294002 is mediated by translocation of BAD and BAX proteins from the cytosol to the mitochondria. Whereas, epidermal growth factor activates a MAPK/ERK/RSK1 module leading to inactivation of BAD via Ser75 phosphorylation, the presence of serum, on the other hand, induces a nonconducive intracellular environment for mitochondrial translocation of dephosphorylated BAD. Taken together, these results indicate that phosphorylation of BAD or inhibition of its translocation to the mitochondria are critical phosphatidylinositol 3-kinase-independent survival pathways in LNCaP cells.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10

Similar content being viewed by others

References

  • Ballif BA, Blenis J . (2001). Cell Growth Differ 12: 397–408.

  • Boldt S, Kolch W . (2004). Curr Pharm Des 10: 1885–1905.

  • Bonni A, Brunet A, West AE, Datta SR, Takasu MA, Greenberg ME . (1999). Science 286: 1358–1362.

  • Carson JP, Kulik G, Weber MJ . (1999). Cancer Res 59: 1449–1453.

  • Chang F, Steelman LS, Shelton JG, Lee JT, Navolanic PM, Blalock WL et al. (2003). Int J Oncol 22: 469–480.

  • Chao DT, Korsmeyer SJ . (1998). Annu Rev Immunol 16: 395–419.

  • Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y et al. (1997). Cell 91: 231–241.

  • Eisenmann KM, VanBrocklin MW, Staffend NA, Kitchen SM, Koo HM . (2003). Cancer Res 63: 8330–8337.

  • Gardai SJ, Hildeman DA, Frankel SK, Whitlock BB, Frasch SC, Borregaard N et al. (2004). J Biol Chem 279: 21085–21095.

  • Green DR, Reed JC . (1998). Science 281: 1309–1312.

  • Horoszewicz JS, Leong SS, Chu TM, Wajsman ZL, Friedman M, Papsidero L et al. (1980). Prog Clin Biol Res 37: 115–132.

  • Horoszewicz JS, Leong SS, Kawinski E, Karr JP, Rosenthal H, Chu TM et al. (1983). Cancer Res 43: 1809–1818.

  • Kajiwara T, Takeuchi T, Ueki T, Moriyama N, Ueki K, Kakizoe T et al. (1999). Int J Urol 6: 520–525.

  • Kooijman R, Coppens A, Hooghe-Peters E . (2003). Cell Signal 15: 1091–1098.

  • Lebedeva I, Rando R, Ojwang J, Cossum P, Stein CA . (2000). Cancer Res 60: 6052–6060.

  • Letai A, Bassik MC, Walensky LD, Sorcinelli MD, Weiler S, Korsmeyer SJ . (2002). Cancer Cell 2: 183–192.

  • Li X, Marani M, Mannucci R, Kinsey B, Andriani F, Nicoletti I et al. (2001). Cancer Res 61: 1699–1706.

  • Linseman DA, Butts BD, Precht TA, Phelps RA, Le SS, Laessig TA et al. (2004). J Neurosci 24: 9993–10002.

  • Lizcano JM, Morrice N, Cohen P . (2000). Biochem J 349: 547–557.

  • Lutz RJ . (2000). Biochem Soc Trans 28: 51–56.

  • Lyons-Darden T, Daaka Y . (2004). J Mol Endocrinol 33: 165–173.

  • Martinou JC, Green DR . (2001). Nat Rev Mol Cell Biol 2: 63–67.

  • McConkey DJ, Greene G, Pettaway CA . (1996). Cancer Res 56: 5594–5599.

  • McDonnell TJ, Troncoso P, Brisbay SM, Logothetis C, Chung LW, Hsieh JT et al. (1992). Cancer Res 52: 6940–6944.

  • McEleny KR, Watson RW, Coffey RN, O'Neill AJ, Fitzpatrick JM . (2002). Prostate 51: 133–140.

  • Murillo H, Huang H, Schmidt LJ, Smith DI, Tindall DJ . (2001). Endocrinology 142: 4795–4805.

  • Nomura T, Mimata H, Takeuchi Y, Yamamoto H, Miyamoto E, Nomura Y . (2003). Urol Res 31: 37–44.

  • Puthalakath H, Strasser A . (2002). Cell Death Differ 9: 505–512.

  • Raffo AJ, Perlman H, Chen MW, Day ML, Streitman JS, Buttyan R . (1995). Cancer Res 55: 4438–4445.

  • Sato S, Fujita N, Tsuruo T . (2004). J Biol Chem 279: 33759–33767.

  • Shahbazi M, Pravica V, Nasreen N, Fakhoury H, Fryer AA, Strange RC et al. (2002). Lancet 359: 397–401.

  • Shi XB, Gumerlock PH, Muenzer JT, deVere White RW . (2001). Cancer Biother Radiopharm 16: 421–429.

  • Shimamura A, Ballif BA, Richards SA, Blenis J . (2000). Curr Biol 10: 127–135.

  • Torring N, Dagnaes-Hansen F, Sorensen BS, Nexo E, Hynes NE . (2003). Prostate 56: 142–149.

  • Yang CC, Lin HP, Chen CS, Yang YT, Tseng PH, Rangnekar VM . (2003). J Biol Chem 278: 25872–25878.

  • Yarden Y . (2001). Eur J Cancer 37 (Suppl 4): S3–S8.

  • Zhang M, Mukherjee N, Bermudez RS, Latham DE, Delaney MA, Zietman AL et al. (2005). Prostate 64: 293–302.

Download references

Acknowledgements

We thank Dr C Brenner for the VDAC antibody, Dr Stanley J Korsmeyer laboratory, Dana-Farber Cancer Institute, Boston, MA, for the pcDNA3-BADwt encoding for murine BAD, Ms Wang Ya for useful technical assistance in the immunofluorescence analysis of activated BAX and Dr S Pervaiz for his help in editing the paper. This work was supported by Grant R-183-000-084-213 from The National Medical Research Council of Singapore to MVC.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to M-V Clément.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chao, O., Clément, MV. Epidermal growth factor and serum activate distinct pathways to inhibit the BH3 only protein BAD in prostate carcinoma LNCaP cells. Oncogene 25, 4458–4469 (2006). https://doi.org/10.1038/sj.onc.1209421

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1209421

Keywords

This article is cited by

Search

Quick links