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 Paper
  • Published:

Association of gp91phox homolog Nox1 with anchorage-independent growth and MAP kinase-activation of transformed human keratinocytes

Oncogene volume 22, pages 6045–6053 (2003)Cite this article

Abstract

Among five members of the NADPH oxidase (Nox) family, Nox1 confers mitogenic properties and is implicated to participate in the process of cell transformation. We have established two phenotypes of carcinogenesis model by ethanol treatment of human gingival keratinocytes immortalized with E6/E7 oncogenes of human papillomavirus type16: immortalized (EPI) nontransformed cells with epithelium-like morphology and more advanced transformed (FIB) cells with spindle fibroblastic-shape morphology. FIB membranes possessed a 63-kDa Nox1 protein at higher levels and exhibited 2.8-fold higher capability for superoxide and hydroxyl radical generation, compared with EPI membranes. Both EPI and FIB cells expressed more abundant Nox1 protein at a proliferating stage than that at a quiescent confluent phase. Immunofluorescence staining with an anti-Nox1 antibody showed that immunoreactive materials were distributed in the whole interior of both types of cells, while they were preferentially localized in the nuclei of FIB cells. Nuclei isolated from EPI and FIB cells contained a 63 kDa-Nox1 protein. Compared with EPI cells, FIB cells expressed elevated levels of Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase proteins. Furthermore, JNK2 was constitutively phosphorylated in FIB cells. Together, our data strongly implicate Nox1 in redox-mediated signaling related to cellular activation of human keratinocytes at a more advanced stage of transformation.

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
Figure 6
Figure 7

Similar content being viewed by others

Abbreviations

DEPMPO:

5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide

DTPA:

dithylenetriamine-pentaacetic acid

EPI:

nontransformed cells with epithelium-like morphology

ERK:

extracellular regulated kinase

FIB:

transformed cells with spindle fibroblastic-shape morphology

FLOAT:

cells with nonadherent free-floating state from FIB cultures

HPV:

human papillomavirus

JNK:

c-Jun N-terminal kinase

KGM:

keratinocyte growth medium

MAP kinase:

mitogen-activated protein kinase

phox :

phagocyte oxidase

•OH:

hydroxyl radical

•OOH:

superoxide radical

SOD:

superoxide dismutase

References

  • Aplin AE, Hogan BP, Tomeu J and Juliano RL . (2002). J. Cell Sci., 115, 2781–2790.

  • Arbiser JL, Petros J, Klafter R, Govindajaran B, McLaughlin ER, Brown LF, Cohen C, Moses M, Kilroy S, Arnold RS and Lambeth JD . (2002). Proc. Natl. Acad. Sci. USA, 99, 715–720.

  • Arnold RS, Shi J, Murad E, Whalen AM, Sun CQ, Polavarapu R, Parthasarathy S, Petros JA and Lambeth JD . (2001). Proc. Natl. Acad. Sci. USA, 98, 5550–5555.

  • Banfi B, Clark RA, Steger K and Krause KH . (2003). J. Biol. Chem., 278, 3510–3513.

  • Banfi B, Maturana A, Jaconi S, Arnaudeau S, Laforge T, Sinha B, Ligeti E, Demaurex N and Krause KH . (2000). Science, 287 (5450), 138–142.

  • Bjerrum OW and Borregaard N . (1989). Eur J. Haematol., 43, 67–77.

  • Chamulitrat W . (1999). Free Radical Biol. Med., 27, 411–421.

  • Chamulitrat W, Schmidt R, Chunglok W, Kohl A and Tomakidi P . (2003). Eur J. Cell. Biol., 82, 313–322.

  • Cheng G, Cao Z, Xu X, van Meir EG and Lambeth JD . (2001). Gene, 269, 131–140.

  • Chiu C, Maddock DA, Zhang Q, Souza KP, Townsend AR and Wan Y . (2001). Int. J. Mol. Med., 8, 251–255.

  • Clark GJ, Cox AD, Graham SM and Der CJ . (1995). Methods Enzymol., Vol. 255, Balch WE, Der CJ and Hall A (eds) Academic Press: New York, pp. 395–412.

    Google Scholar 

  • Elenbaas B and Weinberg RA . (2001). Exp. Cell Res., 264, 169–184.

  • Finkelstein E, Rosen GM, Rauckman EJ and Paxton J . (1979). Mol. Pharmacol., 16, 676–685.

  • Frejaville C, Karoui H, Tuccio B, Le Moigne F, Culcasi M, Pietri S, Lauricella R and Tordo P . (1995). J. Med. Chem., 38, 258–265.

  • Goldman R, Moshonov S and Zor U . (1999). Biochim. Biophys. Acta., 1438, 349–358.

  • Gupta A, Rosenberger SF and Bowden GT . (1999). Carcinogenesis, 20, 2063–2073.

  • Halbert CL, Demers GW and Galloway DA . (1992). J. Virol., 66, 2125–2134.

  • Hei TK, Wu LJ and Piao CQ . (1997). Environ. Health Perspect., 105 (Suppl 5), 1085–1088.

  • Hurlin PJ, Kaur P, Smith PP, Perez-Reyes N, Blanton RA and McDougall JK . (1991). Proc. Natl. Acad. Sci. USA, 88, 570–574.

  • Irani K, Xia Y, Zweier JL, Sollott SJ, Der CJ, Fearon ER, Sundaresan M, Finkel T and Goldschmidt-Clermont PJ . (1997). Science, 275, 1649–1652.

  • Jones RD, Hancock JT and Morice AH . (2000). Free Radical Biol. Med., 29, 416–424.

  • Kuroki T and Huh NH . (1993). Jpn. J. Cancer Res., 84, 1091–1100.

  • Lambeth JD . (2002). Curr. Opin. Hematol., 9, 11–17.

  • Lassegue B, Sorescu D, Szocs K, Yin Q, Akers M, Zhang Y, Grant SL, Lambeth JD and Griendling KK . (2001). Circ. Res., 88, 888–894.

  • Li JM and Shah AM . (2002). J. Biol. Chem., 277, 19952–19960.

  • McCormick JJ and Maher VM . (1988). Mutat. Res., 199, 273–291.

  • Nakamura Y, Ginhart TD, Winterstein D, Tomita I, Seed JL and Colburn NH . (1988). Carcinogenesis, 9, 203–207.

  • Park NH, Gujuluva CN, Baek JH, Cherrick HM, Shin KH and Min BM . (1995). Oncogene, 10, 2145–2153.

  • Plattner R, Gupta S, Khosravi-Far R, Sato KY, Perucho M, Der CJ and Stanbridge EJ . (1999). Oncogene, 18, 1807–1817.

  • Rennefahrt UE, Illert B, Kerkhoff E, Troppmair J and Rapp UR . (2002). J. Biol. Chem., 277, 29510–29518.

  • Rodrigues GA, Park M and Schlessinger J . (1997). EMBO J., 16, 2634–2645.

  • Sashiyama H, Shino Y, Sakao S, Shimada H, Kobayashi S, Ochiai T and Shirasawa H . (2002). Cancer Lett., 177, 21–28.

  • Schimmel M and Bauer G . (2002). Oncogene, 21, 5886–5896.

  • Sen CK, Khanna S, Babior BM, Hunt TK, Ellison EC and Roy S . (2002). J. Biol. Chem., 277, 33284–33290.

  • Sorescu D, Somers MJ, Lassegue B, Grant S, Harrison DG and Griendling KK . (2001). Free Radical Biol. Med., 30, 603–612.

  • Suh YA, Arnold RS, Lassegue B, Shi J, Xu X, Sorescu D, Chung AB, Griendling KK and Lambeth JD . (1999). Nature, 401, 79–82.

  • Teshima S, Kutsumi H, Kawahara T, Kishi K and Rokutan K . (2000). Am. J. Physiol. Gastrointest. Liver Physiol., 279, G1169–G1176.

  • Yang JQ, Li S, Domann FE, Buettner GR and Oberley LW . (1999). Mol. Carcinog., 26, 180–188.

  • Yang X, Hao Y, Pater MM, Tang SC and Pater A . (1998). Mol. Carcinog., 22, 95–101.

  • Yoshida L, Nishida S, Shimoyama T, Kawahara T, Rokutan K and Tsunawaki S . (2002). Biochem. Biophys. Res. Commun., 296, 1322–1328.

Download references

Acknowledgements

We are grateful to Dr D Noethe (University of Heidelberg) for the use of an EPR spectrometer. We thank Dr H Spring of DKFZ for advice on Confocal microscopy. W Chunglok was supported by Thailand Research Fund for graduate studentship. A part of this study was supported by Grants-in-Aid Scientific Research from Japan Society for the Promotion of Science (No. 14370184) and Priority Areas Research from Monbu Kagakusho (No. 14021077) (to K Rokutan). This study was in part supported by Dietmar-Hopp Foundation (to W Stremmel).

Author information

Authors and Affiliations

  1. Department of Applied Tumorvirology, Deutsches Krebsforschungszentrum, Heidelberg, 69120, Germany

    Walee Chamulitrat & Rainer Schmidt

  2. Department of Orthodontics, Dental School, University of Heidelberg, Heidelberg, 69120, Germany

    Pascal Tomakidi

  3. Internal Medicine IV, University Clinic Hospital, University of Heidelberg, Heidelberg, 69115, Germany

    Walee Chamulitrat & Wolfgang Stremmel

  4. Faculty of Medical Technology, Mahidol University, Bangkok, Thailand

    Warangkana Chunglok

  5. Department of Nutrition, School of Medicine, Tokushima University, Tokushima, Japan

    Tsukasa Kawahara & Kazuhito Rokutan

Authors
  1. Walee Chamulitrat
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rainer Schmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pascal Tomakidi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wolfgang Stremmel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Warangkana Chunglok
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tsukasa Kawahara
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kazuhito Rokutan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Walee Chamulitrat.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chamulitrat, W., Schmidt, R., Tomakidi, P. et al. Association of gp91phox homolog Nox1 with anchorage-independent growth and MAP kinase-activation of transformed human keratinocytes. Oncogene 22, 6045–6053 (2003). https://doi.org/10.1038/sj.onc.1206654

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1206654

Keywords

This article is cited by

Search

Advanced search

Quick links