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12-O-tetradecanoylphorbol-1, 3-acetate induces the negative regulation of protein kinase B by protein kinase Cα during gastric cancer cell apoptosis

Cellular & Molecular Biology Letters volume 15, pages 377–394 (2010)Cite this article

Abstract

The PKB signaling pathway is essential for cell survival and the inhibition of apoptosis, but its functional mechanisms have not been fully explored. Previously, we reported that TPA effectively inhibited PKB activity and caused PKB degradation, which was correlated with the repression of PKB phosphorylation at Ser473. In this study, we focus on how PKB is regulated by TPA in gastric cancer cells. One of the TPA targets, PKCα, was found to mediate the inhibition of PKB phosphorylation and degredation caused by TPA. Furthermore, TPA induced the import of PKCα into the nucleus, where PKCα exerted an inhibitory effect on PKB expression and phosphorylation. As a result, cancer cell proliferation was arrested. Our study characterizes a novel function of PKCα in mediating the negative regulation of PKB by TPA, and suggests a potential application in the clinical treatment of gastric cancer.

Abbreviations

BrdU:

5-bromo-2’-deoxyuridine

PI3K:

phosphoinositide-3-OH kinase

PDK1/PDK2:

3-phosphoinositide-dependent kinase

PI:

propidium iodide

PKB:

protein kinase B

PKC:

protein kinase C

TPA:

12-O-tetradecanoylphorbol-1,3-acetate

WGA:

wheat germ agglutinin

References

  1. Brazil, D.P. and Hemmings, B.A. Ten years of protein kinase B signalling: a hard Akt to follow. Trends Biochem. Sci. 26 (2001) 657–664.

    Article  CAS  PubMed  Google Scholar 

  2. Kohn, A.D., Kovacina, K.S. and Roth, R.A. Insulin stimulates the kinase activity of RAC-PK, a pleckstrin homology domain containing ser/thr kinase. EMBO J. 14 (1995) 4288–4295.

    CAS  PubMed  Google Scholar 

  3. Alessi, D.R., Andjelkovic, M., Caudwell, B., Cron, P., Morrice, N., Cohen, P. and Hemmings, B.A. Mechanism of activation of protein kinase B by insulin and IGF-1. EMBO J. 15 (1996) 6541–6551.

    CAS  PubMed  Google Scholar 

  4. Andjelkovic, M., Jakubowicz, T., Corn, P., Ming, X.F., Han, J.W. and Hemmings, B.A. Activation and phosphorylation of a pleckstrin homology domain containing protein kinase (RAC-PK/PKB) promoted by serum and protein phosphatase inhibitors. Proc. Natl. Acad. Sci. USA 93 (1996) 5699–5704.

    Article  CAS  PubMed  Google Scholar 

  5. Vanhaesebroeck, B. and Alessi, D.R. The PI3k-PDK1 connection: more than just a road to PKB. Biochem. J. 346 (2000) 561–576.

    Article  CAS  PubMed  Google Scholar 

  6. Alessi, D.R., James, S.R., Downes, C.P., Holmes, A.B., Gaffney, P.R., Reese, C.B. and Cohen, P. Characterization of a 3-phosphoinositide-dependent protein kinase which phosphorylates and activates protein kinase B alpha. Curr. Biol. 7 (1997) 261–269.

    Article  CAS  PubMed  Google Scholar 

  7. Kroner, C., Eybrechts, K. and Akkerman, J.W. Dual regulation of platelet protein kinase B. J. Biol. Chem. 275 (2000) 27790–27798.

    CAS  PubMed  Google Scholar 

  8. Hill, M.M., Andjelkovic, M., Brazil, D.P., Ferrari, S., Fabbro, D. and Hemmings, B.A. Insulin-stimulated protein kinase B phosphorylation on Ser-473 is independent of its activity and occurs through a staurosporineinsensitive kinase. J. Biol. Chem. 276 (2001) 25643–25646.

    Article  CAS  PubMed  Google Scholar 

  9. Andjelkovic, M., Maira, S.M., Cron, P., Parker, P.J. and Hemmings, B.A. Domain swapping used to investigate the mechanism of protein kinase B regulation by 3-phosphoinositide-dependent protein kinase 1 and Ser473 kinase. Mol. Cell Biol. 19 (1999) 5061–5072.

    CAS  PubMed  Google Scholar 

  10. Chen, D., Fucini, R.V., Olson, A.L., Hemmings, B.A. and Pessin, J.E. Osmotic shock inhibits insulin signaling by maintaining Akt/protein kinase B in an inactive dephosphorylated state. Mol. Cell Biol. 19 (1999) 4684–4694.

    CAS  PubMed  Google Scholar 

  11. Schubert, K.M., Scheid, M.P. and Duronio, V. Ceramide inhibits protein kinase B/Akt by promoting dephosphorylation of serine 473. J. Biol. Chem. 275 (2000) 13330–13335.

    Article  CAS  PubMed  Google Scholar 

  12. Zhang, B. and Xia, C. The expression of protein kinase B in gastric cancer cell apoptosis induced by 12-O-tetradecanoylphorbol-1, 3-acetate. Cell. Mol. Biol. Lett. 14 (2009) 466–480.

    Article  CAS  PubMed  Google Scholar 

  13. Moore, E.D., Ring, M., Scriven, D.R., Smith, V.C., Meloche, R.M. and Buchan, A.M. The role of protein kinase C isozymes in bombesin-stimulated gastrin release from human antral gastrin cells. J. Biol. Chem. 274 (1999) 22493–22501.

    Article  CAS  PubMed  Google Scholar 

  14. Kelly, M.L., Tang, Y., Rosensweig, N., Clejan, S. and Beckman, B.S. Granulocyte-macrophage colony-stimulating factor rescues TF-1 leukemia cells from ionizing radiation-induced apoptosis through a pathway mediated by protein kinase Calpha. Blood 92 (1998) 416–424.

    CAS  PubMed  Google Scholar 

  15. Shao, R.G., Cao, C.X. and Prommier, Y. Activation of PKCalpha downstream from caspases apoptosis induced by 7-hydroxystaurosporine or the topoisomerase inhibitors, camptothecin and etoposide, in human myeloid leukemia HL60 cells. J. Biol. Chem. 272 (1997)31321–31325.

    Article  CAS  PubMed  Google Scholar 

  16. Iioka, Y., Mishima, K., Azuma, N., Tsuchida, A., Takagi, Y., Aoki, T. and Saito, I. Overexpression of protein kinase Cdelta enhances cisplatin-induced cytotoxicity correlated with p53 in gastric cancer cell line. Pathobiology 72 (2005) 152–159.

    Article  CAS  PubMed  Google Scholar 

  17. Endo, K., Kohnoe, S., Tsujita, E., Watanabe, A., Nakashima, H., Baba, H. and Maehara, Y. Modulation of anti-apoptosis by endogenous IAP expression in MKN45 human gastric cancer cells. Anticancer Res. 25 (2005) 2713–2717.

    CAS  PubMed  Google Scholar 

  18. Zhu, G.H., Wong, B.C., Slosberg, E.D., Eggo, M.C., Ching, C.K., Yuen, S.T., Lai, K.C., Soh, J.W., Weinstein, I.B. and Lam, S.K. Overexpression of protein kinase C-beta1 isoenzyme suppresses indomethacin-induced apoptosis in gastric epithelial cells. Gastroenterology 118 (2000) 507–514.

    Article  CAS  PubMed  Google Scholar 

  19. Jiang, X.H., Lam, S.K. Lin, M.C., Jiang, S.H., Kung, H.F. Slosberg E.D., Soh, J.W., Weinstein, I.B. and Wong, B.C. Novel target for induction of apoptosis by cyclo-oxygenase-2 inhibitor SC-236 through a protein kinase C-beta(1)-dependent pathway. Oncogene 21 (2002) 6113–6122.

    Article  CAS  PubMed  Google Scholar 

  20. Li, W., Zhang, J., Flechner, L., Hyun, T., Yam, A., Franke, T.F. and Pierce, J.H. Protein kinase C-alpha overexpression stimulates Akt activity and suppresses apoptosis induced by interleukin 3 withdrawal. Oncogene 18 (1999) 6564–6572.

    Article  CAS  PubMed  Google Scholar 

  21. Doornbos, R.P., Theelen, M., Van Der Hoeven, P.C., Van Blitterswijk, W.J., Verkleij, A.J. and Van Bergen En Henegouwen, P.M. Protein kinase Czeta is a negative regulator of protein kinase B activity. J. Biol. Chem. 274 (1999) 8589–8596.

    Article  CAS  PubMed  Google Scholar 

  22. Kawakami, Y., Nishimoto, H., Kitaura, J., Maeda-Yamamoto, M., Kato, R.M., Littman, D.R., Leitges, M., Rawlings, D.J. and Kawakami, T. Protein kinase C betaII regulates Akt phosphorylation on Ser-473 in a cell type- and stimulus-specific fashion. J. Biol. Chem. 279 (2004) 47720–47725.

    Article  CAS  PubMed  Google Scholar 

  23. Aicher, B., Lerch, M.M., Muller, T., Schilling, J. and Ullrich, A. Cellular redistribution of protein tyrosine phosphatases LAR and PTPsigma by inducible proteolytic processing. J. Biol. Chem. 138 (1997) 681–696.

    CAS  Google Scholar 

  24. Wu, Q., Lin, X.F., Zhang, B., Xie, Z. and Su, W.J. Ubiquitinated or sumoylated retinoic acid receptor alpha determines its characteristic and interacting model with retinoid X receptor alpha in gastric and breast cancer cells. J. Mol. Endocrinol. 32 (2004) 595–613.

    Article  CAS  PubMed  Google Scholar 

  25. Zhao, B.X., Chen, H.Z., Lei, N.Z., Li, G.D., Zhao, W.X., Zhan, Y.Y., Liu, B., Lin, S.C. and Wu, Q. p53 mediates the negative regulation of MDM2 by orphan receptor TR3. EMBO J. 25 (2006) 5703–5715.

    Article  CAS  PubMed  Google Scholar 

  26. Partovian, C., Zhuang, Z., Moodie, K., Lin, M., Ouchi, N., Sessa, W.C., Walsh, K. and Simons, M. PKCalpha activates eNOS and increases arterial blood flow in vivo. Circ. Res. 97 (2005) 482–487.

    Article  CAS  PubMed  Google Scholar 

  27. Chipuk, J.E., Kuwana, T., Bouchier-Hayes, L., Droin, N.M., Newmeyer, D.D., Schuler, M. and Green, D.R. Direct activation of Bax by p53 mediates mitochondrial membrane permeabilization and apoptosis. Science 303 (2004) 1010–1014.

    Article  CAS  PubMed  Google Scholar 

  28. Lu, D., Huang, J. and Basu, A. Deregulation of PKB influences antiapoptotic signaling by PKC in breast cancer cells. Int. J. Oncol. 25 (2004) 671–676.

    CAS  PubMed  Google Scholar 

  29. Wu, Q., Liu, S., Ding, L., Ye, X. and Su, W. PKC alpha translocation from mitochondria to nucleus is closely related to induction of apoptosis in gastric cancer cells. Science in China 45 (2002) 237–244.

    CAS  PubMed  Google Scholar 

  30. Bunn, C.F., Neidig, J.A., Freidinger, K.E., Stankiewicz, T.A., Weaver, B.S., Mcgrew, J. and Allison, L.A. Nucleocytoplasmic shuttling of the thyroid hormone receptor alpha. Mol. Endocrinol. 15 (2001) 512–533.

    Article  CAS  PubMed  Google Scholar 

  31. Hermann, A., Maisel, M., Wegner, F., Liebau, S., Kim, D.W., Gerlach, M., Schwarz, J., Kim, K.S. and Storch, A. Multipotent neural stem cells from the adult tegmentum with dopaminergic potential develop essential properties of functional neurons. Stem Cells 24 (2006) 949–964.

    Article  CAS  PubMed  Google Scholar 

  32. Kolluri, S.K., Bruey-Sedano, N., Cao, X., Lin, B., Lin, F., Han, Y.H., Dawson, M.I. and Zhang, X.K. Mitogenic effect of orphan receptor TR3 and its regulation by MEKK1 in lung cancer cells. Mol. Cell Biol. 23 (2003) 8651–8667.

    Article  CAS  PubMed  Google Scholar 

  33. Balendran, A., Casamayor, A., Deak, M., Paterson, A., Gaffney, P., Currie, R., Downes, P.S., and Alessi, D.R. PDK1 acquires PDK2 activity in the presence of a synthetic peptide derived from the carboxyl terminus of PRK2. Curr. Biol. 9 (1999) 393–404.

    Article  CAS  PubMed  Google Scholar 

  34. Delcommenne, M., Tan, C., Gray, V., Rue, L., Woodgett, J. and Dedhar, S. Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase. Proc. Natl. Acad. Sci. USA 95 (1998) 11211–11216.

    Article  CAS  PubMed  Google Scholar 

  35. Peterson, R.T. and Schreiber, S.L. Kinase phosphorylation: Keeping it all in the family. Curr. Biol. 9 (1999) R521–R524.

    Article  CAS  PubMed  Google Scholar 

  36. Li, L., Sampat, K., Hu, N., Zakari, J. and Yuspa, S.H. Protein kinase C negatively regulates Akt activity and modifies UVC-induced apoptosis in mouse keratinocytes. J. Biol. Chem. 281 (2006) 3237–3243.

    Article  CAS  PubMed  Google Scholar 

  37. Munteanu, A., Taddei, M., Tamburini, I., Bergamini, E., Azzi, A. and Zingg, J.M. Antagonistic effects of oxidized low density lipoprotein and alphatocopherol on CD36 scavenger receptor expression in monocytes: involvement of protein kinase B and peroxisome proliferator-activated receptor-gamma. J. Biol. Chem. 281 (2006) 6489–6497.

    Article  CAS  PubMed  Google Scholar 

  38. Partovian, C. and Simons, M. Regulation of protein kinase B/Akt activity and Ser473 phosphorylation by protein kinase Calpha in endothelial cells. Cell Signal. 16 (2004) 951–957.

    Article  CAS  PubMed  Google Scholar 

  39. Galetic, I., Andjelkovic, M., Meier, R., Brodbeck, D., Park, J. and Hemmings, B.A. Mechanism of protein kinase B activation by insulin/insulin-like growth factor-1 revealed by specific inhibitors of phosphoinositide 3-kinase-significance for diabetes and cancer. Pharmacol. Ther. 82 (1999) 409–425.

    Article  CAS  PubMed  Google Scholar 

  40. Wen, H.C., Huang, W.C., Ali, A., Woodgett, J.R. and Lin, W.W. Negative regulation of phosphatidylinositol 3-kinase and Akt signalling pathway by PKC. Cell. Signal. 15 (2003) 37–45.

    Article  CAS  PubMed  Google Scholar 

  41. Wang, R. and Brattain, M.G. AKT can be activated in the nucleus. Cell. Signal. 18 (2006) 1722–1731.

    Article  CAS  PubMed  Google Scholar 

  42. Newton, A.C. Protein kinase C: structural and spatial regulation by phosphorylation, cofactors, and macromolecular interactions. Chem. Rev. 101 (2001) 2353–2364.

    Article  CAS  PubMed  Google Scholar 

  43. Martelli, A.M., Evangelisti, C., Nyakern, M. and Manzoli, F.A. Nuclear protein kinase C. Biochim. Biophys. Acta 1761 (2006) 542–551.

    CAS  PubMed  Google Scholar 

  44. Eliyahu, E. and Shalgi, R. A role for protein kinase C during rat egg activation. Biol. Reprod. 67 (2002) 189–195.

    Article  CAS  PubMed  Google Scholar 

  45. Carter, C.A., Parham, G.P. and Chambers, T. Cytoskeletal reorganization induced by retinoic acid treatment of human endometrial adenocarcinoma (RL95-2) cells is correlated with alterations in protein kinase C-alpha. Pathobiology 66 (1998) 284–292.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Medical School, Xiamen University, Xiamen, 361005, Fujian Province, China

    Bing Zhang

  2. Zhongshan Hospital, Xiamen University, Xiamen, 361004, Fujian Province, China

    Chun Xia

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  1. Bing Zhang
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  2. Chun Xia
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Correspondence to Chun Xia.

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Zhang, B., Xia, C. 12-O-tetradecanoylphorbol-1, 3-acetate induces the negative regulation of protein kinase B by protein kinase Cα during gastric cancer cell apoptosis. Cell Mol Biol Lett 15, 377–394 (2010). https://doi.org/10.2478/s11658-010-0014-4

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  • DOI: https://doi.org/10.2478/s11658-010-0014-4

Keywords

Cellular & Molecular Biology Letters

ISSN: 1689-1392

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