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:

Protein kinase Cɛ mediates Stat3Ser727 phosphorylation, Stat3-regulated gene expression, and cell invasion in various human cancer cell lines through integration with MAPK cascade (RAF-1, MEK1/2, and ERK1/2)

Oncogene volume 29, pages 3100–3109 (2010)Cite this article

Subjects

Abstract

Protein kinase C epsilon (PKCɛ), a novel calcium-independent PKC isoform, has been shown to be a transforming oncogene. PKCɛ-mediated oncogenic activity is linked to its ability to promote cell survival. However, the mechanisms by which PKCɛ signals cell survival remain elusive. We found that signal transducers and activators of transcription 3 (Stat3), which is constitutively activated in a wide variety of human cancers, is a protein partner of PKCɛ. Stat3 has two conserved amino-acid (Tyr705 and Ser727) residues, which are phosphorylated during Stat3 activation. PKCɛ interacts with Stat3α isoform, which has Ser727, and not with Stat3β isoform, which lacks Ser727. PKCɛ–Stat3 interaction and Stat3Ser727 phosphorylation was initially observed during induction of squamous cell carcinomas and in prostate cancer. Now we present that (1) PKCɛ physically interacts with Stat3α isoform in various human cancer cells: skin melanomas (MeWo and WM266-4), gliomas (T98G and MO59K), bladder (RT-4 and UM-UC-3), colon (Caco-2), lung (H1650), pancreatic (PANC-1), and breast (MCF-7 and MDA:MB-231); (2) inhibition of PKCɛ expression using specific siRNA inhibits Stat3Ser727 phosphorylation, Stat3-DNA binding, Stat3-regulated gene expression as well as cell invasion; and (3) PKCɛ mediates Stat3Ser727 phosphorylation through integration with the MAPK cascade (RAF-1, MEK1/2, and ERK1/2). The results indicate that PKCɛ-mediated Stat3Ser727 phosphorylation is essential for constitutive activation of Stat3 and cell invasion in various human cancers.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

Abbreviations

PCa:

prostate cancer

STAT:

signal transducers and activators of transcription

TRAMP:

transgenic adenocarcinoma of mouse prostate

TPA:

12-O-tetradecanoylphorbol-13-acetate

PKC:

protein kinase C

PS:

phosphatidylserine

DAG:

diacylglycerol

SCC:

squamous cell carcinoma

References

  • Akira S . (2000). Roles of STAT3 defined by tissue-specific gene targeting. Oncogene 19: 2607–2611.

    Article  CAS  PubMed  Google Scholar 

  • Alvarez JV, Febbo PG, Ramaswamy S, Loda M, Richardson A, Frank DA . (2005). Identification of a genetic signature of activated signal transducer and activator of transcription 3 in human tumors. Cancer Res 65: 5054–5062.

    Article  CAS  PubMed  Google Scholar 

  • Aziz MH, Manoharan HT, Church DR, Dreckschmidt NE, Zhong W, Oberley TD et al. (2007a). Protein kinase Cepsilon interacts with signal transducers and activators of transcription 3 (Stat3), phosphorylates Stat3Ser727, and regulates its constitutive activation in prostate cancer. Cancer Res 67: 8828–8838.

    Article  CAS  PubMed  Google Scholar 

  • Aziz MH, Manoharan HT, Sand JM, Verma AK . (2007b). Protein kinase Cepsilon interacts with Stat3 and regulates its activation that is essential for the development of skin cancer. Mol Carcinog 46: 646–653.

    Article  CAS  PubMed  Google Scholar 

  • Aziz MH, Manoharan HT, Verma AK . (2007c). Protein kinase C epsilon, which sensitizes skin to un's UV radiation-induced cutaneous damage and development of squamous cell carcinomas, associates with Stat3. Cancer Res 67: 1385–1394.

    Article  CAS  PubMed  Google Scholar 

  • Bae KM, Wang H, Jiang G, Chen MG, Lu L, Xiao L . (2007). Protein kinase C epsilon is overexpressed in primary human non-small cell lung cancers and functionally required for proliferation of non-small cell lung cancer cells in a p21/Cip1-dependent manner. Cancer Res 67: 6053–6063.

    Article  CAS  PubMed  Google Scholar 

  • Basu A, Sivaprasad U . (2007). Protein kinase Cepsilon makes the life and death decision. Cell Signal 19: 1633–1642.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Burger A, Amemiya Y, Kitching R, Seth AK . (2006). Novel RING E3 ubiquitin ligases in breast cancer. Neoplasia 8: 689–695.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Burger AM, Gao Y, Amemiya Y, Kahn HJ, Kitching R, Yang Y et al. (2005). A novel RING-type ubiquitin ligase breast cancer-associated gene 2 correlates with outcome in invasive breast cancer. Cancer Res 65: 10401–10412.

    Article  CAS  PubMed  Google Scholar 

  • Chan KS, Sano S, Kiguchi K, Anders J, Komazawa N, Takeda J et al. (2004). Disruption of Stat3 reveals a critical role in both the initiation and the promotion stages of epithelial carcinogenesis. J Clin Invest 114: 720–728.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen D, Gould C, Garza R, Gao T, Hampton RY, Newton AC . (2007). Amplitude control of protein kinase C by RINCK, a novel E3 ubiquitin ligase. J Biol Chem 282: 33776–33787.

    Article  CAS  PubMed  Google Scholar 

  • Cornford P, Evans J, Dodson A, Parsons K, Woolfenden A, Neoptolemos J et al. (1999). Protein kinase C isoenzyme patterns characteristically modulated in early prostate cancer. Am J Pathol 154: 137–144.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Decker T, Kovarik P . (2000). Serine phosphorylation of STATs. Oncogene 19: 2628–2637.

    Article  CAS  PubMed  Google Scholar 

  • Griner EM, Kazanietz MG . (2007). Protein kinase C and other diacylglycerol effectors in cancer. Nat Rev Cancer 7: 281–294.

    Article  CAS  PubMed  Google Scholar 

  • Hodge DR, Hurt EM, Farrar WL . (2005). The role of IL-6 and STAT3 in inflammation and cancer. Eur J Cancer 41: 2502–2512.

    Article  CAS  PubMed  Google Scholar 

  • Jain N, Zhang T, Kee WH, Li W, Cao X . (1999). Protein kinase C delta associates with and phosphorylates Stat3 in an interleukin-6-dependent manner. J Biol Chem 274: 24392–24400.

    Article  CAS  PubMed  Google Scholar 

  • Jansen AP, Verwiebe EG, Dreckschmidt NE, Wheeler DL, Oberley TD, Verma AK . (2001). Protein kinase C-epsilon transgenic mice: a unique model for metastatic squamous cell carcinoma. Cancer Res 61: 808–812.

    CAS  PubMed  Google Scholar 

  • Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ . (2009). Cancer statistics, 2009. CA Cancer J Clin 59: 225–249.

    Article  PubMed  Google Scholar 

  • Kazanietz MG . (2000). Eyes wide shut: protein kinase C isozymes are not the only receptors for the phorbol ester tumor promoters. Mol Carcinog 28: 5–11.

    Article  CAS  PubMed  Google Scholar 

  • Klampfer L . (2006). Signal transducers and activators of transcription (STATs): novel targets of chemopreventive and chemotherapeutic drugs. Curr Cancer Drug Targets 6: 107–121.

    Article  CAS  PubMed  Google Scholar 

  • Kobielak A, Fuchs E . (2006). Links between alpha-catenin, NF-kappaB, and squamous cell carcinoma in skin. Proc Natl Acad Sci USA 103: 2322–2327.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kortylewski M, Jove R, Yu H . (2005). Targeting STAT3 affects melanoma on multiple fronts. Cancer Metastasis Rev 24: 315–327.

    Article  CAS  PubMed  Google Scholar 

  • Li L, Shaw PE . (2004). A STAT3 dimer formed by inter-chain disulphide bridging during oxidative stress. Biochem Biophys Res Commun 322: 1005–1011.

    Article  CAS  PubMed  Google Scholar 

  • Mellor H, Parker PJ . (1998). The extended protein kinase C superfamily. Biochem J 332 ( Pt 2): 281–292.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mochly-Rosen D, Kauvar LM . (1998). Modulating protein kinase C signal transduction. Adv Pharmacol 44: 91–145.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Nikitakis NG, Siavash H, Sauk JJ . (2004). Targeting the STAT pathway in head and neck cancer: recent advances and future prospects. Curr Cancer Drug Targets 4: 637–651.

    Article  CAS  PubMed  Google Scholar 

  • Okhrimenko H, Lu W, Xiang C, Hamburger N, Kazimirsky G, Brodie C . (2005). Protein kinase C-epsilon regulates the apoptosis and survival of glioma cells. Cancer Res 65: 7301–7309.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pan Q, Bao LW, Kleer CG, Sabel MS, Griffith KA, Teknos TN et al. (2005). Protein kinase C epsilon is a predictive biomarker of aggressive breast cancer and a validated target for RNA interference anticancer therapy. Cancer Res 65: 8366–8371.

    Article  CAS  PubMed  Google Scholar 

  • Pan Q, Bao LW, Teknos TN, Merajver SD . (2006). Targeted disruption of protein kinase C epsilon reduces cell invasion and motility through inactivation of RhoA and RhoC GTPases in head and neck squamous cell carcinoma. Cancer Res 66: 9379–9384.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Reddig PJ, Dreckschmidt NE, Zou J, Bourguignon SE, Oberley TD, Verma AK . (2000). Transgenic mice overexpressing protein kinase C epsilon in their epidermis exhibit reduced papilloma burden but enhanced carcinoma formation after tumor promotion. Cancer Res 60: 595–602.

    CAS  PubMed  Google Scholar 

  • Rivat C, Rodrigues S, Bruyneel E, Pietu G, Robert A, Redeuilh G et al. (2005). Implication of STAT3 signaling in human colonic cancer cells during intestinal trefoil factor 3 (TFF3)—and vascular endothelial growth factor-mediated cellular invasion and tumor growth. Cancer Res 65: 195–202.

    PubMed  Google Scholar 

  • Stephanou A, Latchman DS . (2005). Opposing actions of STAT-1 and STAT-3. Growth Factors 23: 177–182.

    Article  CAS  PubMed  Google Scholar 

  • Vinkemeier U . (2004). Getting the message across, STAT! Design principles of a molecular signaling circuit. J Cell Biol 167: 197–201.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wheeler DL, Li Y, Verma AK . (2005). Protein kinase C epsilon signals ultraviolet light-induced cutaneous damage and development of squamous cell carcinoma possibly through induction of specific cytokines in a paracrine mechanism. Photochem Photobiol 81: 9–18.

    Article  CAS  PubMed  Google Scholar 

  • Wheeler DL, Martin KE, Ness KJ, Li Y, Dreckschmidt NE, Wartman M et al. (2004). Protein kinase C epsilon is an endogenous photosensitizer that enhances ultraviolet radiation-induced cutaneous damage and development of squamous cell carcinomas. Cancer Res 64: 7756–7765.

    Article  CAS  PubMed  Google Scholar 

  • Wheeler DL, Ness KJ, Oberley TD, Verma AK . (2003). Protein kinase Cepsilon is linked to 12-O-tetradecanoylphorbol-13-acetate-induced tumor necrosis factor-alpha ectodomain shedding and the development of metastatic squamous cell carcinoma in protein kinase Cepsilon transgenic mice. Cancer Res 63: 6547–6555.

    CAS  PubMed  Google Scholar 

  • Wu D, Foreman TL, Gregory CW, McJilton MA, Wescott GG, Ford OH et al. (2002). Protein kinase cepsilon has the potential to advance the recurrence of human prostate cancer. Cancer Res 62: 2423–2429.

    CAS  PubMed  Google Scholar 

  • Wu D, Thakore CU, Wescott GG, McCubrey JA, Terrian DM . (2004). Integrin signaling links protein kinase Cepsilon to the protein kinase B/Akt survival pathway in recurrent prostate cancer cells. Oncogene 23: 8659–8672.

    Article  CAS  PubMed  Google Scholar 

  • Xuan YT, Guo Y, Zhu Y, Wang OL, Rokosh G, Messing RO et al. (2005). Role of the protein kinase C-epsilon-Raf-1-MEK-1/2-p44/42 MAPK signaling cascade in the activation of signal transducers and activators of transcription 1 and 3 and induction of cyclooxygenase-2 after ischemic preconditioning. Circulation 112: 1971–1978.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang Y, Cho YY, Petersen BL, Bode AM, Zhu F, Dong Z . (2003). Ataxia telangiectasia mutated proteins, MAPKs, and RSK2 are involved in the phosphorylation of STAT3. J Biol Chem 278: 12650–12659.

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y, Liu G, Dong Z . (2001). MSK1 and JNKs mediate phosphorylation of STAT3 in UVA-irradiated mouse epidermal JB6 cells. J Biol Chem 276: 42534–42542.

    Article  CAS  PubMed  Google Scholar 

  • Zykova TA, Zhang Y, Zhu F, Bode AM, Dong Z . (2005). The signal transduction networks required for phosphorylation of STAT1 at Ser727 in mouse epidermal JB6 cells in the UVB response and inhibitory mechanisms of tea polyphenols. Carcinogenesis 26: 331–342.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported in parts by DOD Grant W81XWH and NIH Grant CA35368 to AKV.

Author information

Authors and Affiliations

  1. Department of Human Oncology, Wisconsin Institutes for Medical Research, Paul P. Carbone Comprehensive Cancer, School of Medicine and Public Health, University of Wisconsin, Madison, WI, USA

    M H Aziz, B B Hafeez, J M Sand, D B Pierce, S W Aziz, N E Dreckschmidt & A K Verma

  2. Molecular and Environmental Toxicology Center, Wisconsin Institutes of Medical Research, Madison, WI, USA

    J M Sand

Authors
  1. M H Aziz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. B B Hafeez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J M Sand
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D B Pierce
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S W Aziz
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. N E Dreckschmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A K Verma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A K Verma.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Aziz, M., Hafeez, B., Sand, J. et al. Protein kinase Cɛ mediates Stat3Ser727 phosphorylation, Stat3-regulated gene expression, and cell invasion in various human cancer cell lines through integration with MAPK cascade (RAF-1, MEK1/2, and ERK1/2). Oncogene 29, 3100–3109 (2010). https://doi.org/10.1038/onc.2010.63

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2010.63

Keywords

This article is cited by

Search

Advanced search

Quick links