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  • Original Paper
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Growth factors rescue cutaneous melanoma cells from apoptosis induced by knockdown of mutated (V600E) B-RAF

Oncogene volume 24, pages 6292–6302 (2005)Cite this article

Abstract

Constitutive activation of the RAS-RAF-MEK-ERK signaling cascade is a hallmark of cutaneous malignant melanoma. A single activating mutation (c.1799T>A; p.V600E) in the gene encoding the serine/threonine kinase B-RAF occurs in >60% of the tumors. Previous work has shown that knockdown of V600EB-RAF by RNA interference induces a variety of phenotypic changes in cultured melanoma cells, including lower proliferation rates, reduced anchorage-independent growth and apoptosis. Here, we show that the majority of melanomas harboring the V600EB-RAF mutation have retained the wild-type (WT) B-RAF allele, and that these cells can be rescued from the effects of V600EB-RAF knockdown by stimulation with growth factors. Ectopic expression of short hairpin RNAs specifically suppressing V600EB-RAF in melanoma cell lines reduced colony formation by ∼80%. This response could be rescued by basic fibroblast growth factor, hepatocyte growth factor or, to a lesser extent, endothelin-1. Rescue with growth factors was not possible in cell lines lacking WTB-RAF. Single-cell clones with efficient knockdown of V600EB-RAF could be propagated in the presence of basic fibroblast growth factor but underwent apoptosis or senescence-like growth arrest upon withdrawal of this growth factor. The ability of growth factors to modulate the response of V600EB-RAF knockdown in melanoma cells may have both experimental and therapeutic implications.

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References

  • Albino AP, Davis BM and Nanus DM . (1991). Cancer Res., 51, 4815–4820.

  • Bastian BC, LeBoit PE, Hamm H, Bröcker EB and Pinkel D . (1998). Cancer Res., 58, 2170–2175.

  • Berking C, Takemoto R, Satyamoorthy K, Elenitsas R and Herlyn M . (2001). Am. J. Pathol., 158, 943–953.

  • Bielenberg DR, Bucana CD, Sanchez R, Donawho CK, Kripke ML and Fidler IJ . (1998). J. Invest. Dermatol., 111, 864–872.

  • Bridge AJ, Pebernard S, Ducraux A, Nicoulaz AL and Iggo R . (2003). Nat. Genet., 34, 263–264.

  • Brummelkamp TR, Bernards R and Agami R . (2002). Science, 296, 550–553.

  • Collisson EA, De A, Suzuki H, Gambhir SS and Kolodney MS . (2003). Cancer Res., 63, 5669–5673.

  • Davies H, Bignell GR, Cox C, Stephens P, Edkins S, Clegg S, Teague J, Woffendin H, Garnett MJ, Bottomley W, Davis N, Dicks E, Ewing R, Floyd Y, Gray K, Hall S, Hawes R, Hughes J, Kosmidou V, Menzies A, Mould C, Parker A, Stevens C, Watt S, Hooper S, Wilson R, Jayatilake H, Gusterson BA, Cooper C, Shipley J, Hargrave D, Pritchard-Jones K, Maitland N, Chenevix-Trench G, Riggins GJ, Bigner DD, Palmieri G, Cossu A, Flanagan A, Nicholson A, Ho JW, Leung SY, Yuen ST, Weber BL, Seigler HF, Darrow TL, Paterson H, Marais R, Marshall CJ, Wooster R, Stratton MR and Futreal PA . (2002). Nature, 417, 949–954.

  • Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I and Pereira-Smith O . (1995). Proc. Natl. Acad. Sci. USA, 92, 9363–9367.

  • Dong J, Phelps RG, Qiao R, Yao S, Benard O, Ronai Z and Aaronson SA . (2003). Cancer Res., 63, 3883–3885.

  • Easty DJ and Bennett DC . (2000). Melanoma Res., 10, 401–411.

  • Eisinger M, Marko O, Ogata S and Old LJ . (1985). Science, 229, 984–986.

  • Garnett MJ and Marais R . (2004). Cancer Cell, 6, 313–319.

  • Gorden A, Osman I, Gai W, He D, Huang W, Davidson A, Houghton AN, Busam K and Polsky D . (2003). Cancer Res., 63, 3955–3957.

  • Guldberg P, thor Straten P, Birck A, Ahrenkiel V, Kirkin AF and Zeuthen J . (1997). Cancer Res., 57, 3660–3663.

  • Halaban R . (1996). Semin. Oncol., 23, 673–681.

  • Halaban R . (2000). Pigment Cell Res., 13, 4–14.

  • Hingorani SR, Jacobetz MA, Robertson GP, Herlyn M and Tuveson DA . (2003). Cancer Res., 63, 5198–5202.

  • Huang S, Luca M, Gutman M, McConkey DJ, Langley KE, Lyman SD and Bar-Eli M . (1996). Oncogene, 13, 2339–2347.

  • Huser M, Luckett J, Chiloeches A, Mercer K, Iwobi M, Giblett S, Sun XM, Brown J, Marais R and Pritchard C . (2001). EMBO J., 20, 1940–1951.

  • Imokawa G, Kobayasi T and Miyagishi M . (2000). J. Biol. Chem., 275, 33321–33328.

  • Imokawa G, Yada Y and Miyagishi M . (1992). J. Biol. Chem., 267, 24675–24680.

  • Karasarides M, Chiloeches A, Hayward R, Niculescu-Duvaz D, Scanlon I, Friedlos F, Ogilvie L, Hedley D, Martin J, Marshall CJ, Springer CJ and Marais R . (2004). Oncogene, 23, 6292–6298.

  • Kumar R, Angelini S, Czene K, Sauroja I, Hahka-Kemppinen M, Pyrhonen S and Hemminki K . (2003). Clin. Cancer Res., 9, 3362–3368.

  • Li G, Satyamoorthy K, Meier F, Berking C, Bogenrieder T and Herlyn M . (2003). Oncogene, 22, 3162–3171.

  • Li G, Schaider H, Satyamoorthy K, Hanakawa Y, Hashimoto K and Herlyn M . (2001). Oncogene, 20, 8125–8135.

  • Meier F, Nesbit M, Hsu MY, Martin B, Van Belle P, Elder DE, Schaumburg-Lever G, Garbe C, Walz TM, Donatien P, Crombleholme TM and Herlyn M . (2000). Am. J. Pathol., 156, 193–200.

  • Mercer K, Chiloeches A, Huser M, Kiernan M, Marais R and Pritchard C . (2002). Oncogene, 21, 347–355.

  • Nesbit M, Nesbit HK, Bennett J, Andl T, Hsu MY, Dejesus E, McBrian M, Gupta AR, Eck SL and Herlyn M . (1999). Oncogene, 18, 6469–6476.

  • Otsuka T, Takayama H, Sharp R, Celli G, LaRochelle WJ, Bottaro DP, Ellmore N, Vieira W, Owens JW, Anver M and Merlino G . (1998). Cancer Res., 58, 5157–5167.

  • Peyssonnaux C and Eychene A . (2001). Biol. Cell, 93, 53–62.

  • Pollock PM, Harper UL, Hansen KS, Yudt LM, Stark M, Robbins CM, Moses TY, Hostetter G, Wagner U, Kakareka J, Salem G, Pohida T, Heenan P, Duray P, Kallioniemi O, Hayward NK, Trent JM and Meltzer PS . (2003). Nat. Genet., 33, 19–20.

  • Pritchard CA, Samuels ML, Bosch E and McMahon M . (1995). Mol. Cell. Biol., 15, 6430–6442.

  • Satyamoorthy K, Li G, Gerrero MR, Brose MS, Volpe P, Weber BL, Van Belle P, Elder DE and Herlyn M . (2003). Cancer Res., 63, 756–759.

  • Satyamoorthy K, Li G, Vaidya B, Patel D and Herlyn M . (2001). Cancer Res., 61, 7318–7324.

  • Scott PH, Cairns CA, Sutcliffe JE, Alzuherri HM, McLees A, Winter AG and White RJ . (2001). J. Biol. Chem., 276, 1005–1014.

  • Sledz CA, Holko M, de Veer MJ, Silverman RH and Williams BR . (2003). Nat. Cell Biol., 5, 834–839.

  • Smalley KS . (2003). Int. J. Cancer, 104, 527–532.

  • Storm SM, Cleveland JL and Rapp UR . (1990). Oncogene, 5, 345–351.

  • Sumimoto H, Miyagishi M, Miyoshi H, Yamagata S, Shimizu A, Taira K and Kawakami Y . (2004). Oncogene, 23, 6031–6039.

  • Sutcliffe JE, Brown TR, Allison SJ, Scott PH and White RJ . (2000). Mol. Cell. Biol., 20, 9192–9202.

  • Tuveson DA, Weber BL and Herlyn M . (2003). Cancer Cell, 4, 95–98.

  • Wan PT, Garnett MJ, Roe SM, Lee S, Niculescu-Duvaz D, Good VM, Jones CM, Marshall CJ, Springer CJ, Barford D and Marais R . (2004). Cell, 116, 855–867.

  • Wellbrock C, Karasarides M and Marais R . (2004a). Nat. Rev. Mol. Cell. Biol., 5, 875–885.

  • Wellbrock C, Ogilvie L, Hedley D, Karasarides M, Martin J, Niculescu-Duvaz D, Springer CJ and Marais R . (2004b). Cancer Res., 64, 2338–2342.

  • Zhang XD, Borrow JM, Zhang XY, Nguyen T and Hersey P . (2003). Oncogene, 22, 2869–2881.

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Acknowledgements

We thank Vibeke Ahrenkiel for expert technical assistance, Nicole Fehrenbacher, Mikkel Rohde and Jesper Nylandsted Larsen for valuable advice, and Reuven Agami for kindly providing the pSUPER plasmid. This study was supported by grants from the Danish Cancer Society, the Neye Foundation, and the Danish Cancer Research Foundation.

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  1. Institute of Cancer Biology, Danish Cancer Society, Strandboulevarden 49, Copenhagen, DK-2100, Denmark

    Claus Christensen & Per Guldberg

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  1. Claus Christensen
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Correspondence to Per Guldberg.

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Christensen, C., Guldberg, P. Growth factors rescue cutaneous melanoma cells from apoptosis induced by knockdown of mutated (V600E) B-RAF. Oncogene 24, 6292–6302 (2005). https://doi.org/10.1038/sj.onc.1208758

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