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:

Manganese superoxide dismutase deficiency enhances cell turnover via tumor promoter-induced alterations in AP-1 and p53-mediated pathways in a skin cancer model

Abstract

Previous studies in our laboratories demonstrated that overexpression of manganese superoxide dismutase (MnSOD) suppressed both the incidence and multiplicity of papillomas in a DMBA/TPA multi-stage skin carcinogenesis model. The activity of activator protein-1 (AP-1), which is associated with tumor promotion, was reduced in MnSOD transgenic mice overexpressing MnSOD in the skin, suggesting that MnSOD may reduce tumor incidence by suppressing AP-1 activation. In the present study, we report that reduction of MnSOD by heterozygous knockout of the MnSOD gene (Sod2 −/+, MnSOD KO) increased the levels of oxidative damage proteins and the activity of AP-1 following TPA treatment. RNA levels of ornithine decarboxylase (ODC) were also increased, suggesting an increase in cell proliferation in the KO mice. Histological examination confirmed that the number of proliferating cells in DMBA/TPA-treated mouse skin were higher in the KO mice. Interestingly, histological examination also demonstrated greater numbers of apoptotic cells in the KO mice after DMBA/TPA treatment. Evidence of apoptosis, including DNA fragmentation, cytochrome c release from mitochondria, and caspase 3 activation were also observed by biochemical assays of the skin tissues. Apoptosis was associated with an increase in nuclear levels of p53 as determined by Western analysis. Quantitative immunogold ultrastructural analysis confirmed that p53 immunoreactive protein levels were increased to a greater level in the nuclei of epidermal cells from MnSOD KO mice compared to epidermal nuclei from wild type mice similarly treated. Moreover, p53 levels further increased in the mitochondria of DMBA/TPA treated mice, and this increase was much greater in the MnSOD KO than in the wild type mice, suggesting a link between MnSOD deficiency and mitochondrial-mediated apoptosis. Pathological examination reveals no difference in the incidence and frequency of papillomas comparing the KO mice and their wild type littermates. Taken together, these results suggest that: (1) MnSOD deficiency enhanced TPA-induced oxidative stress and AP-1 and p53 levels, consistent with the increase in both proliferation and apoptosis events in the MnSOD KO mice, and (2) increased apoptosis may negate increased proliferation in the MnSOD deficient mice during an early stage of tumor development.

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

Similar content being viewed by others

Abbreviations

The abbreviations used are: 4-HNE:

4-hydroxy-2-nonenal

AP-1:

activator protein-1

DMBA:

7,12-dimethylbenz(α)-anthracene

DMSO:

dimethylsulfoxide

DTT:

dithiothreitol

MnSOD:

manganese superoxide dismutase

EMSA:

electrophoresis mobility shift assay

JNK:

c-Jun terminal kinase

ODC:

ornithine decarboxylase

PBS:

phosphate-buffered saline

PKC:

protein kinase C

PMSF:

phenylmethylfulfonyl fluoride

ROS:

reactive oxygen species

RT–PCR:

reverse transcription-PCR

TBS:

Tris-buffered saline

TPA:

12-O-tetradecanoylphorbol-13-acetate

References

  • Agarwal R, Khan SG, Athar M, Zaidi SIA, Bickers DR, Mukhtar H . 1993 Mol. Carcinog. 8: 290–298

  • Amstad PA, Liu H, Ichimiya M, Berezesky IK, Trump BF . 1997 Carcinogenesis 18: 479–484

  • Bowden GT, Finch J, Domann F, Krieg P . 1995 Molecular mechanisms involved in skin tumor initiation, promotion and progression Mukhtar H (ed) Skin cancer: mechanisms and human relevance Boca Raton, FL: CRC Press, Inc pp 99–111

    Google Scholar 

  • Bernstein LR, Ben-Ari ET, Simek SL, Colburn NH . 1991 Environ. Health Perspect. 93: 111–119

  • Church SL, Grant JW, Ridnour LA, Oberley LW, Swanson PE, Meltzer PS, Trent JM . 1993 Proc. Natl. Acad. Sci. USA 90: 3113–3117

  • Das KC, Guo XL, White CW . 1998 J. Biol. Chem. 273: 34639–34645

  • Donahue RJ, Razmara M, Hoek JB, Knodsen TB . 2001 FASEB J. 15: 635–644

  • Drane P, Bravard A, Bouvard V, May E . 2001 Oncogene 20: 430–439

  • Du C, Fang M, Li Y, Li L, Wang X . 2000 Cell 102: 33–42

  • Epperly MW, Sikora C, Defilippi S, Bray J, Koe G, Liggitt D, Luketich JD, Greenberger JS . 2000 Int. J. Cancer 90: 128–137

  • Fischer SM, Jasheway DW, Klann RC, Butler AP, Patrick KE, Baldwin JK, Cameron GS . 1989 Cancer Res. 49: 6693–6699

  • Goldstein JC, Waterhouse NJ, Juin P, Evan GI, Green DR . 2000 Nat. Cell Biol. 2: 156–162

  • Gonzalez-Zulueta M, Ensz LM, Mukhina G, Lebovitz RM, Zwacka RM, Engelhardt JF, Oberley LW, Dawson VL, Dawson TM . 1998 J. Neurosci. 18: 2040–2055

  • Gopalakrishna R, Anderson WB . 1989 Proc. Natl. Acad. Sci. USA 86: 6758–6762

  • Guidot DM, Rapine JE, Kitlowski AD, Flores SC, Nelson SK, Wright RM, McCord JM . 1995 J. Clin. Invest. 96: 1131–1136

  • Hohmeier HE, Thigpen A, Tran VV, Davis R, Newgard CB . 1998 J. Clin. Invest. 101: 1811–1820

  • Karin M . 1995 J. Biol. Chem. 270: 16483–16486

  • Keller JN, Kindy M, Holtsberg FW, St. Clair DK, Yen H-C, Mattson MP . 1998 J. Neurosci. 18: 687–697

  • Khajuria A, Thusu N, Zutshi U, Bedi KL . 1998 Mol. Cell. Biochem. 189: 113–118

  • Kinscherf R, Claus R, Wagner M, Gehrke C, Kamencic H, Hou D, Nauen O, Schmiedt W, Kovacs G, Pill J, Metz J, Deigner H-P . 1998 FASEB J. 12: 461–467

  • Kiningham KK, Oberley TD, Lin S, Mattingly CA, St. Clair DK . 1999 FASEB J. 13: 1601–1610

  • Lam EW, Zwacka R, Engelhardt JF, Davidson BL, Domann FE, Yan T, Oberley LW . 1997 Cancer Res. 57: 5550–5556

  • Lam EW, Zwacka R, Seftor EA, Nieva DR, Davidson BL, Engelhardt JF, Hendrix MJ, Oberley LW . 1999 Free Radic. Biol. Med. 27: 572–579

  • Li JJ, Oberley LW, St. Clair DK, Ridnour LA, Oberley TD . 1995 Oncogene 10: 1989–2000

  • Li N, Oberley TD, Oberley LW, Zhong W . 1998 Prostate 35: 221–233

  • Li P-F, Dietz R, von Harsdorf Rudiger . 1999 EMBO J. 18: 6027–6036

  • Liu R, Oberley TD, Oberley LW . 1997 Hum. Gene Ther. 8: 585–595

  • Majima HJ, Oberley TD, Furakawa K, Mattson MP, Yen HC, Szweda LI, St. Clair DK . 1998 J. Biol. Chem. 273: 8217–8224

  • Marchenko ND, Zaika AI, Moll UM . 2000 J. Biol. Chem. 275: 16202–16212

  • McGarrity TJ, Peiffer LP, Granville DJ, Carthy CM, Levy JG, Khandelwal M, Hunt DW . 2001 Cancer Lett. 163: 33–41

  • McSwine-Kennick RL, McKeegan EM, Johnson MD, Morin MJ . 1991 J. Biol. Chem. 266: 15135–15143

  • Miyashita T, Reed JC . 1995 Cell 80: 293–299

  • Moll UM, Zaika A . 2001 FEBS Lett. 493: 65–69

  • Nakamura Y, Kamamoto N, Ohto Y, Torikai K, Murakami A, Ohigashi H . 1999 Cancer Lett. 140: 37–45

  • Nishizuka Y . 1988 Nature 334: 661–665

  • Oda K, Arakawa H, Tanaka T, Matsuda K, Tanikawa C, Mori T, Nishimori H, Tamai K, Tokino T, Nakamura Y, Taya Y . 2000 Cell 102: 849–862

  • Pence B, Reiners JJ . 1987 Cancer Res. 47: 6388–6392

  • Polyak K, Xia Y, Zweier JL, Kinzler KW, Vogelstein B . 1997 Nature 389: 300–305

  • Porter AG . 1999 Trends Cell. Biol. 10: 394–401

  • Reiners JJ, Pence BC, Barcus MCS, Cantu A . 1987 Cancer Res. 47: 1775–1779

  • Reiners JJ, Thai G, Rupp T, Cantu AR . 1991 Carcinogenesis 12: 2337–2343

  • Remmen HV, Salvador C, Yang H, Huang TT, Epstein CJ, Richardson A . 1999 Arch. Biochem. Biophys. 363: 91–97

  • Ruggeri BA, Klein-Szanto AJP . 1995 Alterations in the p53 and Rb-1 tumor suppression genes in carcinogen-induced murine skin models of squamous cell neoplasias. Mukhtar H (ed) Skin Cancer: mechanisms and human relevance Boca Raton, FL: CRC Press, Inc pp 339–347

    Google Scholar 

  • Safford SE, Oberley TD, Urano M, St. Clair DK . 1994 Cancer Res. 54: 4261–4265

  • Soengas MS, Alarcon RM, Yoshida H, Giaccia AJ, Hakem R, Mak TW, Lowe SW . 1999 Science 284: 156–159

  • Spitz DR, Oberley LW . 1989 Anal. Biochem. 179: 8–18

  • St. Clair DK, Oberley TD, Ho YS . 1991 FEBS Lett. 293: 199–202

  • Stadtman ER . 1992 Science 257: 1220–1224

  • Susin SA, Lorenzo HK, Zamzami N, Marzo I, Snow BE, Brothers GM, Mangion J, Jacotot E, Costantini P, Loeffler M, Larochette N, Goodlett DR, Aebersold R, Siderovski DP, Penninger JM, Kroemer G . 1999 Nature 397: 441–446

  • Tokino T, Nakamura Y . 2000 Crit. Rev. Oncol. Hematol. 33: 1–6

  • Urano M, Kuroda M, Reynolds R, Oberley TD, St. Clair DK . 1995 Cancer Res. 55: 2490–2493

  • Verhagen AM, Ekert PG, Pakusch M, Silke J, Connolly LM, Reid GE, Moritz RL, Simpson RJ, Vaux DL . 2000 Cell 102: 43–53

  • Verma AJ, Pong RC, Erickson D . 1986 Cancer Res. 46: 6149–6155

  • Weisiger RA, Fridovich I . 1973 J. Biol. Chem. 248: 4793–4796

  • Wong GHW, Elwell JH, Oberley LW, Goeddel D . 1989 Cell 58: 923–931

  • Xu Y, Krishnan A, Wan XS, Majima H, Yeh CC, Ludewig G, Kasarskis EJ, St. Clair DK . 1999 Oncogene 18: 93–102

  • Yan T, Oberley LW, Zhong W, St. Clair DK . 1996 Cancer Res. 56: 2864–2871

  • Young MR, Li JJ, Rincon M, Flavell RA, Sathyanarayana BK, Hunziker R, Colburn N . 1999 Proc. Natl. Acad. Sci. USA 96: 9827–9832

  • Zhao Y, Kiningham KK, Lin S, St. Clair DK . 2001a Antioxidant and Redox Signaling 3: 375–386

  • Zhao Y, Xue Y, Oberley TD, Kiningham KK, Lin SM, Yen HC, Majima H, Hines J, St. Clair DK . 2001b Cancer Res. 61: 6082–6088

  • Zhong W, Oberley LW, Oberley TD, St. Clair DK . 1997 Oncogene 14: 481–490

Download references

Acknowledgements

The authors wish to thank Dr Judy Hines, Cathleen Hsu and Marcie Cole for their technical assistance. This work was supported by NIH grants CA 73599, CA 73599-S1, and AG-08938. Luksana Chaiswing is partially supported by the Thailand Research Fund under the Golden Jubilee Program.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Daret St. Clair.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhao, Y., Oberley, T., Chaiswing, L. et al. Manganese superoxide dismutase deficiency enhances cell turnover via tumor promoter-induced alterations in AP-1 and p53-mediated pathways in a skin cancer model. Oncogene 21, 3836–3846 (2002). https://doi.org/10.1038/sj.onc.1205477

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1205477

Keywords

This article is cited by

Search

Quick links