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  • Original Paper
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MGSA/GRO-mediated melanocyte transformation involves induction of Ras expression

Oncogene volume 19, pages 4647–4659 (2000)Cite this article

Abstract

The MGSA/GRO protein is endogenously expressed in almost 70% of the melanoma cell lines and tumors, but not in normal melanocytes. We have previously demonstrated that over-expression of human MGSA/GROα, β or γ in immortalized murine melanocytes (melan-a cells) enables these cells to form tumors in SCID and nude mice. To examine the possibility that the MGSA/GRO effect on melanocyte transformation requires expression of other genes, differential display was performed. One of the mRNA's identified in the screen as overexpressed in MGSA/GRO transformed melan-a clones was the newly described M-Ras or R-Ras3 gene, a member of the Ras gene superfamily. Over-expression of MGSA/GRO upregulates M-Ras expression at both the mRNA and protein levels, and this induction requires an intact glutamine-leucine-arginine (ELR)-motif in the MGSA/GRO protein. Western blot examination of Ras expression revealed that K- and N-Ras proteins are also elevated in MGSA/GRO-expressing melan-a clones, leading to an overall increase in the amount of activated Ras. MGSA/GRO-expressing melan-a clones exhibited enhanced AP-1 activity. The effects of MGSA/GRO on AP-1 activation could be mimicked by over-expression of wild-type M-Ras or a constitutively activated M-Ras mutant in control melan-a cells as monitored by an AP-1-luciferase reporter, while expression of a dominant negative M-Ras blocked AP-1-luciferase activity in MGSA/GRO-transformed melan-a clones. In the in vitro transformation assay, over-expression of M-Ras mimicked the effects of MGSA/GRO by inducing cellular transformation in control melan-a cells, while over-expression of dominant negative M-Ras in MGSA/GROα-expressing melan-a-6 cells blocked transformation. These data suggest that MGSA/GRO-mediated transformation requires Ras activation in melanocytes.

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Abbreviations

CXC chemokine:

chemokine with the first two conserved cysteine residues separated by an intervening amino acid

CMV:

cytomegalovirus

DMEM:

Dulbecco's modified Eagle's medium

IL-8:

interleukin-8

MGSA/GRO:

melanoma growth-stimulatory activity/growth-regulated protein

AP-1:

activation protein-1

RT-PCR:

reverse transcription-polymerase chain reaction

SV-40:

simian virus-40

EMSA:

electrophoresis mobility shift assay

and MMLV:

moloney murine virus

ELR motif:

glutamine-leucine-arginine tripeptide sequence near the N-terminus of MGSA/GRO proteins

References

  • Arenberg DA, Polverini PJ, Kunkel S, Shanafelt A, Hesselgesser J, Horuk R and Strieter M. . 1997 J. Leukocyte Biol. 62: 554–562.

  • Balentien E, Mufson BE, Shattuck RL, Derynck R and Richmond A. . 1991 Oncogene 6: 1115–1124.

  • Bennett DC, Cooper PJ and Hart IR. . 1987 Int. J. Cancer 39: 414–418.

  • Cao Y, Chen C, Weatherbee JA, Tsang M and Folkman J. . 1995 J. Exp. Med. 182: 2069–2077.

  • Chin L, Tam A, Pomerantz J, Wong M, Holash J, Bardeesy N, Shen Q, O'Hagan R, Pantginis J, Zhou H, Horner JW, Cordon-Cardo C, Yancopoulos GD and DePinho RA. . 1999 Nature 400: 468–472.

  • Della Rocca GJ, van Biesen T, Daaka Y, Luttrell DK, Luttrell LM and Lefkowitz RJ. . 1997 J. Biol. Chem. 272: 19125–19132.

  • Domann FE Jr, Levy JP, Finch JS and Bowden GT. . 1994 Mol. Carcinog. 9: 61–66.

  • Fujimoto J, Ichigo S, Hori M, Morishita S and Tamaya T. . 1995 Mol. Biol. 55: 25–33.

  • Geiser T, Dewald B, Ehrengruber MU, Clark-Lewis I and Baggiolini M. . 1993 J. Biol. Chem. 268: 15419–15424.

  • George DL, Glick B, Trusko S and Freeman N. . 1986 Proc. Natl. Acad. Sci. USA 83: 1651–1655.

  • Haghnegahdar H, Du J, Wang D, Strieter RM, Burdick MD, Nanney LB, Cardwell N, Luan J, Shattuck-Brandt R and Richmond A. . 2000 J. Leuk. Biol. 67: 53–62.

  • Haskill S, Peace A, Morris J, Sporn SA, Anisowicz A, Lee SW, Smith T, Martin G, Ralph P and Sager R. . 1990 Proc. Natl. Acad. Sci. USA 87: 7732–7736.

  • Hawes BE, Luttrell LM, van Biesen T and Lefkowitz RJ. . 1996 J. Biol. Chem. 271: 12133–12136.

  • Hébert CA, Vitangcol RV and Baker JB. . 1991 J. Biol. Chem. 266: 18989–18994.

  • Hesselgesser J, Chitnis CE, Miller LH, Yansura DG, Simmons LC, Fairbrother WJ, Kotts C, Wirth C, Gillece-Castro BL and Horuk R. . 1995 J. Biol. Chem. 270: 11472–11476.

  • Huang HS, Nagane M, Klingbeil CK, Lin H, Nishikawa R, Ji XD, Huang CM, Gill GN, Wiley HS and Cavenee WK. . 1997 J. Biol. Chem. 272: 2927–2935.

  • Husain Z, Yang QM and Biswas D. . 1990 Arch. Dermatol. 126: 324–330.

  • Iida N and Grotendorst GR. . 1990 Mol. Cell. Biol. 10: 5596–5599.

  • Johnson R, Spiegelman BM, Hanahan D and Wisdom R. . 1996 Mol. Cell. Biol. 18: 4504–4511.

  • Kimmelman A, Tolkacheva T, Lorenzi MV, Osada M and Chan AM. . 1997 Oncogene 15: 2675–2685.

  • Knall C, Worthen GS and Johnson GL. . 1997 Proc. Natl. Acad. Sci. USA 94: 3052–3057.

  • Kolibaba KS and Druker BJ. . 1997 Biochim. Biophys. Acta. 1333: F217–F248.

  • Kurdowska A, Cohen AB, Carr FK, Stevens MD, Miller EJ, Mullenbach G and Tekamp-Olson P. . 1994 Cytokine 6: 124–134.

  • Limatola C, Mileo AM, Giovannelli A, Vacca F, Ciotti MT, Mercanti D, Santoni A and Eusebi F. . 1999 J. Biol. Chem. 274: 36537–36543.

  • Lloyd AC, Yancheva N and Wasylyk B. . 1991 Nature 352: 635–638.

  • Lopez-Ilasaca M, Crespo P, Pellici PG, Gutkind JS and Wetzker R. . 1997 Science 275: 394–397.

  • Louahed J, Grasso L, De Smet C, Van Roost E, Wildmann C, Nicolaides NC, Levitt RC and Renauld JC. . 1999 Blood 94: 1701–1710.

  • Luan J, Shattuck-Brandt R, Haghnegahdar H, Owen JD, Strieter R, Burdick M, Nirodi C, Beauchamp D, Johnson KN and Richmond A. . 1997 J. Leuk. Biol. 62: 588–597.

  • Mangues R, Seidman I, Gordon JW and Pellicer A. . 1992 Oncogene 7: 2073–2076.

  • Matsumoto K, Asano T and Endo T. . 1997 Oncogene 15: 2409–2417.

  • Mattei S, Colombo MP, Melani C, Silvani A, Parmiani G and Herlyn M. . 1994 Int. J. Cancer 56: 853–857.

  • McKay IA, Marshall CJ, Cales C and Hall A. . 1986 EMBO J. 5: 2617–2621.

  • Neptune ER and Bourne HR. . 1997 Proc. Natl. Acad. Sci. USA 94: 14489–14494.

  • Owen JD, Strieter R, Burdick M, Haghnegahdar H, Nanney L, Shattuck-Brandt R and Richmond A. . 1997 Int. J. Cancer 73: 94–103.

  • Quilliam LA, Castro AF, Rogers-Graham KS, Martin CB, Der CJ and Bi C. . 1999 J. Biol. Chem. 274: 23850–23857.

  • Rennekampff HO, Hansbrough JF, Woods V, Jr; Dore C, Kiessig V and Schroder JM. . 1997 Arch. Dermatol. Res. 289: 204–212.

  • Richmond A and Thomas HG. . 1986 J. Cell. Physiol. 129: 375–384.

  • Richmond A and Thomas HG. . 1988 J. Cell. Biochem. 36: 185–198.

  • Rodeck U, Becker D and Herlyn M. . 1991 Cancer Cells 3: 308–311.

  • Rosenkilde MM and Schwartz TW. . 2000 Mol. Pharmacol. 57: 602–609.

  • Rutberg SE, Lee EJ, Hansen LH, Glick AB and Yuspa SH. . 1997 Mol. Carcinog. 20: 88–98.

  • Sadhu C, Masinovsky B and Staunton DE. . 1998 J. Immunol. 160: 5622–5628.

  • Saez E, Rutberg SE and Mueller E. . 1995 Cell 82: 721–732.

  • Shattuck RL, Wood LD, Jaffe GJ and Richmond A. . 1994 Mol. Cell. Biol. 14: 791–802.

  • Shattuck-Brandt RL, Wood LD and Richmond A. . 1997 DNA Sequence 7: 379–386.

  • Shattuck-Brandt RL and Richmond A. . 1997 Cancer Res. 57: 3032–3039.

  • Shyamala V and Khoja H. . 1998 Biochemistry 37: 15918–15924.

  • Smith JJ, Derynck R and Korc M. . 1987 Proc. Natl. Acad. Sci. USA 84: 7567–7570.

  • Taylor SJ and Shalloway D. . 1996 Curr. Biol. 6: 1621–1627.

  • van Dam H, Huguier S, Kooistra K, Baguet J, Vial E, Vandereb AJ, Herrlich P, Angel P and Castellazzi M. . 1998 Genes Dev. 12: 1227–1239.

  • Vojtek AB and Der CJ. . 1998 J. Biol. Chem. 273: 19925–19928.

  • Westaway D, Papkoff J, Moscovici C and Varmus HE. . 1986 EMBO. J. 5: 301–309.

  • Winzen R, Kracht M, Ritter B, Wilhelm A, Chen C-YA, Shyu A-B, Muller M, Gaestel M, Resch K and Holtmann H. . 1999 EMBO J. 18: 4969–4980.

  • Wood LD and Richmond A. . 1995 J. Biol. Chem. 270: 30619–30626.

  • Zhang B, Shih JW, Wear DJ, Tsai S and Lo SC. . 1997 Proc. Sco. Exp. Biol. Med 214: 359–366.

  • Yang W, Wang D and Richmond A. . 1999 J. Biol. Chem. 274: 11328–11333.

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Acknowledgements

We are indebted to the NIH for support: CA56704 (A Richmond); Vanderbilt Cancer Center CA 68485, Department of Veterans Affairs Merit Award and Career Scientist Award (A Richmond), and to James Owen and Eddy Balentien for developing the MGSA/GRO expressing melan-a clones. We are also indebted to Ben Johnston and Amy Pruitt for excellent technical assistance.

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Authors and Affiliations

  1. Department of Cell Biology, Vanderbilt University School of Medicine, Nashville, 37232, Tennessee, TN, USA

    Dingzhi Wang, Wei Yang, Jianguo Du, Madhav N Devalaraja, Peng Liang & Ann Richmond

  2. VA Medical Center, Nashville, 37232, Tennessee, TN, USA

    Ann Richmond

  3. Department of Biology, Chiba University, Japan

    Ken Matsumoto, Keisuke Tsubakimoto & Takeshi Endo

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Wang, D., Yang, W., Du, J. et al. MGSA/GRO-mediated melanocyte transformation involves induction of Ras expression. Oncogene 19, 4647–4659 (2000). https://doi.org/10.1038/sj.onc.1203820

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