Growth factors rescue cutaneous melanoma cells from apoptosis induced by knockdown of mutated (V 600 E) B-RAF

Oncogene. 2005 Sep 15;24(41):6292-302. doi: 10.1038/sj.onc.1208758.


Constitutive activation of the RAS-RAF-MEK-ERK signaling cascade is a hallmark of cutaneous malignant melanoma. A single activating mutation (c.1799 T>A; p.V 600 E) in the gene encoding the serine/threonine kinase B-RAF occurs in >60% of the tumors. Previous work has shown that knockdown of (V 600 E)B-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 (V 600 E)B-RAF mutation have retained the wild-type (WT) B-RAF allele, and that these cells can be rescued from the effects of (V 600 E)B-RAF knockdown by stimulation with growth factors. Ectopic expression of short hairpin RNAs specifically suppressing (V 600 E)B-RAF in melanoma cell lines reduced colony formation by approximately 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 (WT)B-RAF. Single-cell clones with efficient knockdown of (V 600 E)B-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 (V 600 E)B-RAF knockdown in melanoma cells may have both experimental and therapeutic implications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Apoptosis / physiology*
  • Base Sequence
  • Cell Line, Tumor
  • DNA Primers
  • Growth Substances / physiology*
  • Humans
  • Melanoma / genetics
  • Melanoma / pathology*
  • Mutation*
  • Proto-Oncogene Proteins B-raf / genetics*
  • Skin Neoplasms / genetics
  • Skin Neoplasms / pathology*


  • DNA Primers
  • Growth Substances
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf